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Heat shock protein and the skin

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Abstract

Summary Heat shock proteins are of fundamental importance in cutaneous biology, from protection against UV-induced damage to wound healing and repair. Heat shock proteins have important regulatory roles in the control of apoptosis, regulation of steroid aporeceptors, kinases, and other protein remodelling events. They are also implicated in the control of cell growth, and as such, are potential targets for cancer diagnosis and treatment. Currently, emphasis is being placed on the potential use of these proteins in the prevention and treatment of disease. Therapeutic manipulation of these proteins may ultimately lead to novel treatments for diseases as diverse as melanoma to epidermolysis bullosa.

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... The major stress-induced member of Hsp proteins, Hsp70, has been reported to effectively protect various cell types from cell death [16,17]. Based on previous investigations with regard to Hsp, particularly Hsp70, it was shown that Hsp70 is expressed in keratinocytes not only in vitro but also in vivo [18][19][20]. Moreover, it was demonstrated by Trautinger et al. [21] that high expression of Hsp70 protects human skin against UVR-induced epidermal damage. ...
... The extraction of epidermal tissue to quantify Hsp70 was performed by Extraction Reagent (19) provided with Hsp70 enzyme-linked immunosorbent assay (ELISA) kit containing protease inhibitor cocktail (Sigma). After 30 min incubation at 4°C, the homogenate was centrifugated (15 min, 14,000 g). ...
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Melatonin, a lipophilic compound synthesized and released from the pineal gland, effectively acts against ultraviolet radiation (UVR), one of the main inducers of epidermal damage, skin cancer, inflammation, and DNA photodamage. One of the common known stress protein induced by UVR is heat shock protein 70 (Hsp70), highly expressed in human keratinocytes, providing cellular resistance to such stressors. Here, using human full-thickness skin and normal human epidermal keratinocytes (NHEK), we investigated the interaction of melatonin and Hsp70 towards UVR-induced inflammatory and apoptotic responses. The following observations were made: (i) UVR up-regulated Hsp70 gene expression in human epidermis while melatonin significantly inverted this effect, (ii) similar patterns of regulation were observed within Hsp70 protein level, (iii) mechanistic studies involving silencing of Hsp70 RNA (Hsp70 siRNA) showed prominent decrease of IκB-α (an inhibitor of NF-κB) and enhanced gene expression of pro-inflammatory cytokines (IL-1β, IL-6, Casp-1) and pro-apoptotic protein (Casp-3) in NHEK. Parallel investigation using melatonin (10–3 M) significantly inverted these responses regardless depletion of Hsp70 RNA suggesting a compensatory action of this compound in the defense mechanisms. Our findings combined with data reported so far thus enrich existing knowledge about the potent anti-apoptotic and anti-inflammatory action of melatonin.This article is protected by copyright. All rights reserved.
... We believe that the abnormal response to various environmental stresses in hypertension is genetically-determined and that genes induced by heat or other stressors, modify the development of hypertension. Heat shock proteins (HSP) include 5 major protein families in mammalian cells, denoted as HSP110, HSP90, HSP70, HSP27 and HSP8.5 according to their approximate molecular weight [19]. The genes encoding HSP are highly conserved evolutionally and play important roles in normal cell function [19][20][21][22][23][24]. ...
... Heat shock proteins (HSP) include 5 major protein families in mammalian cells, denoted as HSP110, HSP90, HSP70, HSP27 and HSP8.5 according to their approximate molecular weight [19]. The genes encoding HSP are highly conserved evolutionally and play important roles in normal cell function [19][20][21][22][23][24]. HSP27, HSPT0 and HSP104 have been found to be involved in the regulation of thermotolerance [25][26][27][28]. ...
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Previous studies in our laboratory have demonstrated that the thermosensitivity locus cosegregates with blood pressure and that the elevated expression and restriction fragment length polymorphism of HSP70 gene are associated with hypertension. Cell protection against environmental stressors such as heat and chemicals is often accompanied by up-regulated expression of a wide spectrum of heat shock genes(HSP). To further investigate the interrelation between HSP expression and blood pressure regulation, we employed an effective method of cloning 2 potential hypertension-related HSPs. Synthetic oligonucleotides corresponding either to a highly-conserved region of the known HSP family or a repetitive sequence in the protein encoding gene were used as target primers for polymerase chain reaction (PCR). cDNA prepared from heat-stressed and non-stressed vascular smooth muscle cells (VSMC) of Brown Norway rats (BN.lx) and spontaneously hypertensive rats (SHRp) respectively served as template in the reaction. The PCR products were subsequently analyzed in a single-stranded conformational polymorphism (SSCP) electrophoresing system. Differential gene expression in BN. lx and SHRp was seen on autoradiographs of SSCP gel by comparing the migration patterns of PCR-amplified DNA fragments. Using this technique, we also found that HSP27 and a new member of the large HSP gene family were differentially expressed in BN.1x and SHRp VSMC.
... HSP70 expression in the skin is increased by various stresses, including heat [62,63]. As a protection mechanism, HSP expression in keratinocytes is also increased by UV irradiation [63][64][65][66][67]. ...
... HSP70 expression in the skin is increased by various stresses, including heat [62,63]. As a protection mechanism, HSP expression in keratinocytes is also increased by UV irradiation [63][64][65][66][67]. Moreover, HSP70 overexpression in keratinocytes and melanocytes protected cells against injury from UV in in vitro and in vivo models [62,[67][68][69][70][71]. ...
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Excess melanin deposition in the skin causes cosmetic problems. HSP70 upregulation decreases microphthalmia-associated transcription factor (MITF) expression, which eventually decreases tyrosinase activity and melanogenesis. Ultraviolet (UV) radiation upregulates p53, which increases the melanocortin receptor (MC1R) and MITF. Furthermore, HSP70 decreases p53 and radiofrequency irradiation (RF) increases HSP70. We evaluated whether RF increased HSP70 and decreased p53, consequently decreasing the MITF/tyrosinase pathway and melanogenesis in UV-B radiated animal skin. Various RF combinations with 50, 100, and 150 ms and 5, 10, and 15 W were performed on the UV-B radiated mouse skin every 2 d for 28 d. When RF was performed with 100 ms/10 W, melanin deposition, evaluated by Fontana–Masson staining, decreased without skin crust formation in the UV-B radiated skin. Thus, we evaluated the effect of RF on decreasing melanogenesis in the HEMn and UV-B radiated skin at a setting of 100 ms/10 W. HSP70 expression was decreased in the UV-B radiated skin but was increased by RF. The expression of p53, MC1R, and MITF increased in the UV-B radiated skin but was decreased by RF. The expression of p53, MC1R, and MITF increased in the α-MSH treated HEMn but was decreased by RF. The decreasing effects of RF on p53, MC1R, CREB and MITF were higher than those of HSP70-overexpressed HEMn. The decreasing effect of RF on p53, MC1R, CREB, and MITF disappeared in the HSP70-silenced HEMn. MC1R, CREB, and MITF were not significantly decreased by the p53 inhibitor in α-MSH treated HEMn. RF induced a greater decrease in MC1R, CREB, and MITF than the p53 inhibitor. Therefore, RF may have decreased melanin synthesis by increasing HSP70 and decreasing p53, thus decreasing MC1R/CREB/MITF and tyrosinase activity.
... These genes are polymorphic, with some variants potentially accounting for a change in function and susceptibility to stress tolerance [14,15]. HSP70 gene polymorphisms were found to be risk factors in several human disorders15161718. They might play an important role in susceptibility to and/or progression of diabetic nephropathy. ...
Article
HSPs (heat-shock proteins) are molecular chaperones synthesized under stress conditions, and are involved in renal cell survival and matrix remodelling in acute and chronic renal diseases. In the present study, we investigated whether the HSP70 gene polymorphisms affect susceptibility to DN (diabetic nephropathy) in patients with T2DM (Type 2 diabetes mellitus). The study group consisted of 452 patients with nephropathy. Two control subgroups involved 340 healthy individuals and 132 patients with T2DM lasting > or =10 years who were free of nephropathy. Subjects were genotyped for the HSP70-1 +190 G/C and -110 A/C, HSP70-2 +1267 A/G and HSP70-hom +2437 T/C polymorphisms by PCR, followed by digestion with restriction endonucleases. There were no statistically significant differences in genotype distribution between patients with T2DM with DN and controls for the HSP70-hom polymorphism. Significant differences were observed for HSP70-1 and HSP70-2 polymorphisms. CC homozygotes of the -110 and +190 HSP70-1 polymorphisms were more frequent in patients with T2DM with DN compared with healthy controls (22 compared with 6% and 15 compared with 6.5% respectively; P<0.01). The OR (odds ratio) for the risk allele was 2.17 [95% CI (confidence interval), 1.73-2.72] for the -110 A/C and 1.74 (95% CI, 1.40-2.15) for +190 G/C polymorphisms. A strong association with DN was found for the +1267 HSP70-2 polymorphism. The GG genotype and the G allele were associated with DN, with the OR for the G allele being 4.77 (95% CI, 3.81-5.96). All GG homozygotes in the patient group had higher LDL (low-density lipoprotein)-cholesterol levels than AA homozygotes (P<0.01), suggesting that the observed effect might be associated with this cardiovascular risk factor. These patients progressed faster to end-stage renal failure than those with other genotypes. In conclusion, our results indicate that the HSP70-1 and HSP70-2 polymorphisms are associated with renal complications in T2DM and may be useful in identifying patients with increased risk of DN.
... It has neuroprotective and cardioprotective functions mediated through enhanced manganese superoxide dismutase activity. This is associated with mitochondrial protection and prevention of apoptosis (Gusev et al., 2005; Benn et al., 2002; Morris, 2002). HSP27 seems to play important roles in signal transduction. ...
Article
Heat shock proteins (HSPs) are molecular chaperones involved in protein folding, assembly and transport, and which play critical roles in the regulation of cell growth, survival and differentiation. We set out to test the hypothesis that HSP27 protein is expressed in the human testes and its expression varies with the state of spermatogenesis. HSP27 expression was examined in 30 human testicular biopsy specimens (normal spermatogenesis, maturation arrest and Sertoli cell only syndrome, 10 cases each) using immunofluorescent methods. The biopsies were obtained from patients undergoing investigations for infertility. The seminiferous epithelium of the human testes showing normal spermatogenesis had a cell type-specific expression of HSP27. HSP27 expression was strong in the cytoplasm of the Sertoli cells, spermatogonia, and Leydig cells. Alternatively, the expression was moderate in the spermatocytes, weak in the spermatids and absent in the spermatozoa. In testes showing maturation arrest, HSP27 expression was strong in the Sertoli cells, weak in the spermatogonia, and spermatocytes. It was absent in the spermatids and Leydig cells. In Sertoli cell only syndrome, HSP27 expression was strong in the Sertoli cells and absent in the Leydig cells. We report for the first time the expression patterns of HSP27 in the human testes and show differential expression during normal spermatogenesis, indicating a possible role in this process. The altered expression of this protein in testes showing abnormal spermatogenesis may be related to the pathogenesis of male infertility.
... can be induced by a variety of adverse environmental and pathophysiological conditions, including hyperthermia , sulfhydryl reagents, or transition metal ions (Lindquist 1986; Lindquist and Craig 1988; Macario and Conway de Macario 2005). Overexpression of Hsp70 by transfection of hsp70 cDNA has been reported to enhance myocardial tolerance to ischemia-reperfusion injury in rats (Suzuki et al 1997Suzuki et al , 2002 Marber et al 1995) and in mice (Plumier et al 1995Plumier et al , 1997 Trost et al 1998 ). ...
Article
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Heat shock protein 70 (Hsp70) comprises proteins that have been reported to protect cells, tissues, and organisms against damage from a wide variety of stressful stimuli; however, little is known about whether Hsp70 protects against DNA damage. In this study, we investigated the relationship between Hsp70 expression and the levels of ultraviolet C (UVC)–induced DNA damage in A549 cells with normal, inhibited, and overexpressed Hsp70 levels. Hsp70 expression was inhibited by treatment with quercetin or overexpressed by transfection of plasmids harboring the hsp70 gene. The level of DNA damage was assessed by the comet assay. The results showed that the levels of DNA damage (shown as the percentage of comet cells) in A549 cells increased in all cells after exposure to an incident dose of 0, 10, 20, 40, and 80 J/m 2 whether Hsp70 was inhibited or overexpressed. This response was dose dependent: a protection against UVC-induced DNA damage in cells with overexpressed Hsp70 was observed at UVC dose 20 J/ m 2 with a maximum at 40 J/m 2 when compared with cells with normal Hsp70 levels and in quercetin-treated cells. This differential protection disappeared at 80 J/m 2. These results suggest that overexpressed Hsp70 might play a role in protecting A549 cells from DNA damage caused by UVC irradiation, with a threshold of protection from at UVC irradiation induced DNA damage by Hsp70. The detailed mechanism how Hsp70 is involved in DNA damage and possible DNA repair warrants further investigation.
... Heat Shock Proteins (HSPs), which are widely distributed molecular chaperones and exude a protective response when a cell is under stress, are important regulators of apoptosis [45]. Hsp27, hsp47, hsp60, hsp70, and hsp90 are known to be constitutively expressed in healthy skin; nevertheless, their expression is elevated in stressful situations such as the wound environment [46,47]. It is critical to comprehend how these proteins are expressed in keloid scars because, by Totan et al. [48], collagen synthesis and the expression of hsp27 and hsp47 are intricately linked. ...
Article
Keloids are benign tumors that grow as a result of excessive collagen release from overexpressed fibroblasts. Keloids and hypertrophic scars are distinguished by the fact that keloids develop beyond the site of the original lesion, but hypertrophic scars do not. It is still unclear why this mechanism operates, mainly when aberrant scarring occurs. Despite several treatments' availability, the keloids' recurrence rate remains high. Here, we summarize recent narrative reviews, systematic reviews, and meta-analyses to give a general overview of the condition, its underlying causes, and available therapies for keloids. To undertake a comprehensive investigation of the disease, over 100 publications were reviewed utilizing Google Scholar and Pubmed. We also shed light on using phytochemicals as a natural alternative to prevent keloid scarring, which occurs when the body responds abnormally to external injuries by producing scar tissue. We also summarize the available current treatments.
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Atopic dermatitis (AD) is a chronic inflammatory skin disease and sometimes is a tough challenge for physicians. We previously reported that in Th2 environment, the production and secretion of thymic stromal lymphopoietin (TSLP) from human keratinocytes was inhibited by recombinant heat shock protein 70 (rHSP70). The present study assessed the therapeutic effectiveness of rHSP70 in a mouse model of AD. An experimental model of AD was reproduced by systemic sensitization and local epicutaneous challenge with ovalbumin (OVA). Treatment of rHSP70 was performed by subcutaneous administration. The levels of OVA‐specific IgE, as well as cytokines, were detected by ELISA. Skin samples from patch areas were also taken for histologic examination. Injection of rHSP70 improved the histologic picture by reducing the thickness of epidermis and allergic inflammation. Skin sonography revealed rHSP70 ameliorated skin remodeling. rHSP70 also significantly decreased the protein expression of TSLP of skin from patch areas. Furthermore, in ex vivo studies also showed group of rHSP70 treatment decreased IL‐13, RANTES, MIP‐1β and increased IFN‐γ secreted from splenocytes stimulated with OVA. The rHSP70 intervention in the mouse model of AD reduced the skin expression of TSLP and attenuated the clinical appearance of OVA‐induced AD mice. The effect was achieved by suppressed Th2 immune response in injected skin tissue and enhanced systemic Th1 immune response. These results suggest that rHSP70 have potential as a promising protein for the treatment of AD.
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Accumulation of molecular damage and increased molecular heterogeneity are hallmarks of photoaged skin and pathogenesis of human cutaneous disease. Growing evidence demonstrates the ability of molecular chaperone proteins and of pharmacologic chaperones to decrease the environmental stress and ameliorate the oxidation stress-related and glycation disease phenotypes, suggesting that the field of chaperone therapy might hold novel treatments for skin diseases and aging. In this review, we examine the evidence suggesting a role for molecular chaperone proteins in the skin and their inducer and protecting agents: pharmacologic chaperone imidazole dipeptide-based agents (carcinine and related compounds) in cosmetics and dermatology. Furthermore, we discuss the use of chaperone therapy for the treatment of skin photoaging diseases and other skin pathologies that have a component of increased glycation and/or free radical-induced oxidation in their genesis. We examine biologic activities of molecular and pharmacologic chaperones, including strategies for identifying potential chaperone compounds and for experimentally demonstrating chaperone activity in in vitro and in vivo models of human skin disease. This allows the protein to function and traffic to the appropriate location in the skin, thereby increasing protein activity and cellular function and reducing stress on skin cells. The benefits of imidazole dipeptide antioxidants with transglycating activity (such as carcinine) in skin care are that they help protect and repair cell membrane damage and help retain youthful, younger-looking skin. All skin types will benefit from daily, topical application of pharmacologic chaperone antioxidants, anti-irritants, in combination with water-binding protein agents that work to mimic the structure and function of healthy skin. General strategies are presented addressing ground techniques to improve absorption of usually active chaperone proteins and dipeptide compounds, include encapsulation into hydrophobic carriers, a combination with penetration enhancers, active electrical transport, or chemical modification to increase hydrophobicity.
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Background: Heat shock proteins (HSPs), a group of heat stress proteins, are characterized by highly conserved properties. Malignant transformation is a cellular stress, and the expression of HSPs may be affected during this process. Heat shock protein 105 (HSP105) is a protective protein that has long been observed in many cancer types, but little attention has been given to cutaneous squamous cell carcinoma (CSCC). As such, the objectives of this study were to observe the expression of HSP105 on CSCC and evaluate its correlation with clinicopathological characteristics. Methods: This retrospective study enrolled 60 patients with CSCC. The patients' clinical data, including sex, age, tumor location, tumor type, and degree of pathological differentiation, were collected. The expression of HSP105 was measured by Western blot and immunohistochemical staining. Results: HSP105 expression was decreased in CSCC (HSCORE=0.65 (0.30, 1.98)) compared with normal skin (HSCORE=2.20 (1.50, 2.80)) (P<0.001). These results were consistent with the Western blot analysis. HSP105 immunostaining of Bowen disease (HSCORE=1.28 (1.08, 2.40)) revealed higher expression than in verrucous carcinoma (HSCORE=0.30 (0.23, 0.85)), keratoacanthoma (HSCORE=0.53 (0.29, 0.93)) and acantholytic squamous cell carcinoma (HSCORE=0.53 (0.41, 0.68) (P<0.01)). Poorly differentiated CSCC showed significantly higher expression of HSP105. Conclusion: Our study reveals for the first time that the expression of HSP105 is decreased in CSCC. We suggest that the molecular mechanisms underlying the differential expression of HSP deserve a more rigorous future study, the results of which might explain its role in carcinogenesis and its potential as a target for selective tumor therapy.
Article
The expression of heat shock proteins (HSPs), particularly HSP70, provides resistance to stressors. We recently reported that ultraviolet (UV)-induced melanin production and skin damage were suppressed in transgenic mice expressing HSP70 and that an extract of Eupatorium lindleyanum induces the expression of HSP70 in cells. Here we report the purification of eupalinolide A and B (EA and EB) from E. lindleyanum, and describe their actions as HSP-inducers. EA and EB both induced the expression of HSP70 in cells at concentrations that did not significantly affect cell viability. Treatment of cells with EA or EB activated heat shock factor 1 (HSF1), while the artificial suppression of HSF1 expression diminished the EA- or EB-mediated induction of HSP70 expression. Furthermore, EB inhibited the interaction between HSF1 and HSP90, which is known to inhibit the activity of HSF1. These findings suggest that EA and EB induce the expression of HSP70 via the activation of HSF1 by inhibiting the interaction between HSF1 and HSP90. EA and EB both induced the expression of HSP70 synergistically with other stressors. Furthermore, pre-treatment of cells with EA or EB suppressed melanin production and stressor-induced apoptosis. These effects were suppressed by the artificial suppression of HSP70 expression. In vivo, the percutaneous administration of EB induced the expression of HSP70 and suppressed UVB radiation-induced damage, inflammatory responses and melanin production in the skin. These results suggest that EA and EB could be beneficial for use in cosmetics and medicines as a consequence of their inhibitory action on UV-induced skin damage and melanin production.
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Clinically, keloids are defined as scars that invade adjacent healthy tissue and are caused by tissue injury; however, the clear distinction relating keloids to wound healing or to cancer remains elusive. This study profiled protein extracts from the regions of keloid tissues to determine the activities within these different zones, and to help underpin the activities apparent within different regions of the disease. Two-dimensional gel electrophoresis, liquid chromatography mass spectrometer (LCMS) and a Mascot online database search were employed for comparative proteomic analysis between four sites in keloidal scars (KS). Out of 400 spots identified in all gels, 21 were unique to given KS sites. These were identified using LCMS and the results showed the presence of mitochondrial and structural proteins in the margin, while the top contained keratin II. Heat shock protein was the only protein present in the internal normal control and margin, while the external normal contain keratin II and the voltage-dependent anion-selective channel protein, annexin A6, plus glial fibrillary acidic protein. This work is novel since it has identified differentially expressed proteins across the tested keloidal scar sites. The presence of mitochondrial-associated proteins at the margins suggests that this is the most active part of the scar.
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We propose a new dual treatment strategy for venous ulcer, consisting in simultaneous modulation of matrix metallo-proteinases (MMPs) and heat shock proteins (HSPs) in the wound. One treatment method which can efficiently modu-late both these substances is based on the application of dual-frequency ultrasound (LDM). This strategy was checked in a pilot study on 10 patients with chronic venous ulcers and demonstrated excellent healing rate.
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This experiment compared secondary hair follicles (SFs) in Tibetan cashmere goats from two different steppes that were at different altitudes and had different temperatures. Twenty-four 2-year-old goats were studied. Twelve goats were from Rikaze in Tibet which is at an altitude of above 5000 m with an average temperature of 0 °C. The other 12 studied goats were from Huan County of Gansu Province which is around 2000 m above sea level with an average temperature of 9.2 °C. The structural features of SFs were assessed using light microscopy and transmission electron microscopy. The presence of HIF-1a, HIF-2a, HIF-3a, HSP27, and HOXC13 proteins was studied using immunohistochemistry and immunofluorescence. Light and electron microscopy revealed that the SFs of the Tibetan cashmere goats that lived in the Rikaze Steppe were in the proanagen stage in May. However, the SFs of the goats from the lower warmer Huan County were in the anagen stage at the same time. Immunohistochemistry revealed intense immunostaining for HIF-1a protein in the inner root sheath (IRS) and hair shaft (HS); immunostaining against HIF-2a in the outer root sheath (ORS) and IRS; HIF-3a protein immunostaining in the ORS; HSP27 immunostaining in the ORS, IRS, and HS; and HOXC13 immunostaining in the ORS and HS. HIF-1a protein expression in the IRS and HS was higher than the expression in the ORS (p < 0.05) while the expression of HIF-2a protein was higher in the ORS and IRS than the HS (p < 0.05). The expression of HIF-3a protein was higher in the ORS than in the IRS (p < 0.05). Expression of HOXC13 protein was higher in the ORS than in the IRS and HS (p < 0.05). Immunostaining of HIF-1a, HIF-2a, and HSP27 protein was significantly higher in SFs from cashmere goats from Rikaze than in goats from Huan (p < 0.05). In contrast, HOX13 protein immunostaining was significantly higher in cashmere goats from Huan than from Rikaze (p < 0.05). Significant differences were observed in the SFs of cashmere goats from two locations that differ in altitude and temperature. This suggests the differences in the secondary hair follicles could be due to the hypoxia and lower temperatures experienced by the goats in Rikaze. These results are useful in understanding how altitude and temperature influence SF development. Hair produced by the SFs are used for down fiber. Therefore, understanding of the factors that influence SF development will allow the production and harvest of these valuable fibers to be maximized.
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In oral cavity, the tongue is the most common site prone to development of squamous cell carcinoma (SCC). Considering malignant transformation as a cellular stress, the expression of heat shock proteins (HSPs) may be affected in this process. In this study we assessed the expression of HSP105 and HSP27 as two of the most interested stress proteins and investigated their relationship with grade and stage of the tongue SCC. Fifty-six specimens including 31 early and 25 advanced tongue SCC were gathered. All specimens were graded histologically from I to III. Sixteen sections of normal oral mucosa were used as control group. The cellularity and intensity of HSP105 and HSP27 expression were studied immunohistochemically in both case and control groups. Results were expressed by histochemical score (HSCORE). Significant differences were observed between expression of HSPs and stage of the disease. From early to advanced stage, the expression of HSP105 and HSP27 increased and decreased, respectively. There was no relationship between histological grade of lesion and HSCORE of HSP105 expression (P=0.5), although, HSP27 expression had reverse relationship with the SCC histological grade. HSP27 and HSP105 may be indicated for prognostic purposes in evaluation of tongue SCC. HSP 27 may be used for more accurate microscopic grading of tongue SCC. Increased expression of HSP105 in advanced stage may lead to using this protein for immunotherapy of tongue SCC.
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Molecular chaperone, heat shock proteins (HSPs), stabilizes intracellular processes of cells under stress. Little is known about the role of molecular chaperone proteins in the skin pathology, rejuvenation and wound healing, or whether their expression is altered by environmental and physiological stress to the skin or systemic disease. The focus of this study was to examine the role of molecular chaperone proteins in the skin’s local response to wounding, skin ageing and a range of skin diseases. Free radicals, one form of insult, induce or contribute to adverse effects on the skin, including erythema, oedema, wrinkling, photoaging, inflammation, autoimmune reactions, hypersensitivity, keratinization abnormalities, preneoplastic lesions and skin cancer. A unified view of the molecular and cellular pathogenesis of the skin age-related pathology conditions has led to the search for molecular and chemical chaperones that can slow, arrest or revert disease progression. Specific alpha-crystallin domains and pharmacological imidazole-containing dipeptide chaperone molecules are now emerging that link our biophysical insights with developed skin therapeutic techniques. In this article, we discuss the molecular nature of the stress signals, the mechanisms that underlie activation of the heat shock response, the role of molecular chaperone proteins as skin protective molecules, and strategies for pharmacologically active chaperone molecules and their imidazole-containing dipeptide inducers as regulators of the skin stress response. We discuss how impairment in protein hydration may cause ultrastructural, mechanical and biochemical changes in structural proteins in the aged skin. We have pioneered the molecular chaperone protein activated therapeutic or cosmetic platform to enable simultaneous analysis of water-binding and structuring characteristics for biology of skin ageing and skin disease-related pathways. This cutting-edge technology has improved the way that proteins hydrate in photoaged skin. The mechanisms of skin diseases, ageing, cellular, and signalling pathways mediated by targeting with molecular chaperone protein(s) in patented formulations with imidazole containing dipeptide (N-acetylcarnosine, carcinine, carnosine) are also discussed within this review. Les chaperonnes moléculaires, protéines des chocs thermiques (Heat Shock Proteins), stabilisent les processus intracellulaires lors de stress. Peu de choses sont Connues à propos du rôle des protéines chaperonnes moléculaires dans les pathologies cutanées, le rajeunissement et la cicatrisation, ou à propos des modifications de leur expression lors de stress cutanés environnementaux et physiologiques, ou lors de maladies systémiques. Cette étude est centrée sur l’examen du rôle des protéines chaperonne moléculaires dans la réponse locale cutanée lors de blessure, lors du vieillissement cutané et lors de maladies cutanées. Les radicaux libres, une des formes d’agression, incitent ou contribuent à des effets cutanés délétères comme l’érythème, l’œdème, l’apparition de rides, le photo vieillissement, l’inflammation, des réactions auto immunes, des phénomènes d’hypersensibilité, des anomalies de kératinisation, des lésions pré néoplasiques et le cancer cutané. Une vue globale de la pathogénie moléculaire et cellulaire des conditions pathologiques liées à l’âge ont menéà des recherches sur les chaperonnes qui peuvent ralentir, arrêter ou inverser la progression de maladie. Des domaines alpha-cristallin spécifique et contenant un dipeptide pharmacologique imidazole au sein des chaperonnes sont mis en évidence ce qui relie nos données biophysiques avec les techniques de thérapeutiques cutanées. Dans cet article nous discutons la nature moléculaire des signaux de stress, les mécanismes qui provoquent l’activation de la réponse aux chocs thermiques, le rôle de protéines chaperonnes comme molécules protectrices de la peau et des stratégies pour obtenir des chaperonnes pharmacologiquement actives et leurs inducteurs contenant le dipeptide imidazole comme régulateurs de la réponse au stress cutané. Nous discutons comment la diminution de l’hydratation de la protéine peut engendrer des changements d’ultrastructure, de mécanique et de biochimie des protéines structurelles dans la peau âgée. Nous avons ouvert la voie par la protéine chaperonne d’une plateforme thérapeutique ou cosmétique pour permettre l’analyse simultanée des caractéristiques d’hydratation et de structure pour la biologie, le vieillissement cutané et les voies de pathologie. Cette technologie de pointe a amélioré la connaissance de l’hydratation protéique dans la peau photo vieillie. Les mécanismes de pathologies, de vieillissement, de fonctionnement cellulaire et les voies de signalisation obtenues par des moyens de ciblage avec les protéines chaperonnes moléculaires contenant le dipeptide imidazole (N-acetylcarnosine, carcinine, carnosine) dans des formulations brevetées sont également discutés dans cet article.
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Objective Skin‐brightening agents prevent melanogenesis and reduce melanin production. However, a lower melanin content leads to weaker protection against sunlight. In this study, we evaluated the effect of lysophosphatidylcholine (LPC) and its commercial‐grade product, Lysofix Dry™ (LD), on heat shock protein 70 (HSP70) expression in epidermal cells and their anti‐skin photoaging effect against ultraviolet B (UVB) and blue light. Methods The HSP70 induction was detected using ELISA. To confirm the inhibition of melanin synthesis by LPC or LD, the melanin content assay and gene expression were analyzed. Cell viability was assessed to verify whether LPC or LD prevents photo‐induced skin damage. The split‐face test was performed to confirm skin brightening effect of LD. Cream formulation with 2% of LD and placebo were used for 8 weeks and skin brightness (L) was measured with chromameter (CR‐400, Konica Minolta). Results LPC and LD induced HSP70 expression in epidermal cells. LPC and LD effectively suppressed melanogenesis provoked by α‐MSH in B16 cells. They also inhibited the mRNA transcription of MITF and tyrosinase under blue light irradiation. LD increased the viability of B16 and HaCaT cells after UVB and blue light irradiation in vitro. The cream containing 2% LD increased ΔL by 1.7 after 8 weeks of use, whereas the placebo led to an increase of 0.7. Conclusion LPC and LD were effective in suppressing melanogenesis and enhancing cell viability under UVB and blue light via HSP70 expression. Thus, they can be considered as potent skin brightening agents with protective effects against skin photoaging.
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Skin hyperpigmentation disorders due to abnormal melanin production induced by ultraviolet (UV) irradiation are both a clinical and cosmetic problem. UV irradiation stimulates melanin production in melanocytes by increasing intracellular cAMP. Expression of heat shock proteins (HSPs), especially HSP70, is induced by various stressors, including UV irradiation, to provide cellular resistance to such stressors. In this study we examined the effect of expression of HSP70 on melanin production both in vitro and in vivo. 3-Isobutyl-1-methylxanthine (IBMX), a cAMP-elevating agent, stimulated melanin production in cultured mouse melanoma cells, and this stimulation was suppressed in cells overexpressing HSP70. IBMX-dependent transcriptional activation of the tyrosinase gene was also suppressed in HSP70-overexpressing cells. Expression of microphthalmia-associated transcription factor (MITF), which positively regulates transcription of the tyrosinase gene, was up-regulated by IBMX; however, this up-regulation was not suppressed in HSP70-overexpressing cells. On the other hand, immunoprecipitation and immunostaining analyses revealed a physical interaction between and co-localization of MITF and HSP70, respectively. Furthermore, the transcription of tyrosinase gene in nuclear extract was inhibited by HSP70. In vivo, UV irradiation of wild-type mice increased the amount of melanin in the basal layer of the epidermis, and this increase was suppressed in transgenic mice expressing HSP70. This study provides the first evidence of an inhibitory effect of HSP70 on melanin production both in vitro and in vivo. This effect seems to be mediated by modulation of MITF activity through a direct interaction between HSP70 and MITF.
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Irradiation with UV light, especially UVB, causes epidermal damage via the induction of apoptosis, inflammatory responses, and DNA damage. Various stressors, including UV light, induce heat shock proteins (HSPs) and the induction, particularly that of HSP70, provides cellular resistance to such stressors. The anti-inflammatory activity of HSP70, such as its inhibition of nuclear factor kappa B (NF-kappaB), was recently revealed. These in vitro results suggest that HSP70 protects against UVB-induced epidermal damage. Here we tested this idea by using transgenic mice expressing HSP70 and cultured keratinocytes. Irradiation of wild-type mice with UVB caused epidermal damage such as induction of apoptosis, which was suppressed in transgenic mice expressing HSP70. UVB-induced apoptosis in cultured keratinocytes was suppressed by overexpression of HSP70. Irradiation of wild-type mice with UVB decreased the cutaneous level of IkappaB-alpha (an inhibitor of NF-kappaB) and increased the infiltration of leukocytes and levels of pro-inflammatory cytokines and chemokines in the epidermis. These inflammatory responses were suppressed in transgenic mice expressing HSP70. In vitro, the overexpression of HSP70 suppressed the expression of pro-inflammatory cytokines and chemokines and increased the level of IkappaB-alpha in keratinocytes irradiated with UVB. UVB induced an increase in cutaneous levels of cyclobutane pyrimidine dimers and 8-hydroxy-2'-deoxyguanosine, both of which were suppressed in transgenic mice expressing HSP70. This study provides genetic evidence that HSP70 protects the epidermis from UVB-induced radiation damage. The findings here also suggest that the protective action of HSP70 is mediated by anti-apoptotic, anti-inflammatory, and anti-DNA damage effects.
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The heat shock proteins (HSPs), originally identified as heat-inducible gene products, are a highly conserved family of proteins that respond to a wide variety of stress. Although HSPs are among the most abundant intracellular proteins, they are expressed at low levels under normal physiological conditions, and show marked induction in response to various stressors. HSPs function primarily as molecular chaperones, facilitating the folding of other cellular proteins, preventing protein aggregation, or targeting improperly folded proteins to specific pathways for degradation. By modulating inflammation, wound debris clearance, cell proliferation, migration and collagen synthesis, HSPs are essential for normal wound healing of the skin. In this review, our goal is to discuss the role and clinical implications of HSP with respect to skin wound healing and diabetes. The numerous defects in the function of HSPs associated with diabetes could contribute to the commonly observed complications and delayed wound healing in diabetics. Several physical, pharmacological and genetic approaches may be considered to address HSP-directed therapies both in the laboratory and in the clinics.
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The expression of heat shock proteins (Hsp) expression is induced in all cells by exposure to heat and other environmental stress and Hsp can protect cells from damage through further exposure. Hsp are highly conserved and it is likely that they are essential for survival in a potentially harmful environment. Most Hsp are molecular chaperones sensing unfolded proteins and mediating their re-folding, transport, and interaction. In human epidermis Hsp are associated with differentiation, photoprotection, and skin disease. Recent research has mainly focused on the 27kD and 72kD Hsp that are constitutively expressed in keratinocytes. Cell death induced by ultraviolet radiation (UV) can be inhibited by previous heat shock and UV itself can induce Hsp experimentally. Regulation of Hsp can be pharmacologically modified and topical and systemic inducers and inhibitors of Hsp expression are under development. Whether phototherapy exerts its clinical efficacy by modulation of Hsp has not been sufficiently studied. The UV-wavelength ranges, -intensities and -doses that are required to interfere with the heat shock response in the skin still remain to be elucidated.
Article
Proteomic analysis of murine skin has shown that a variety of heat shock proteins (HSPs) are constitutively expressed in the skin. Using murine allergic contact hypersensitivity as a model, we investigated the role of two heat shock proteins, HSP27 and HSP70, in the induction of cutaneous cell-mediated immune responses. Immunohistochemical examination of skin specimens showed that HSP27 was present in the epidermis and HSP70 was present in both the epidermis and dermis. Inhibition of HSP27 and HSP70 produced a reduction in the 1-fluoro-2,4-dinitrobenzene contact hypersensitivity response and resulted in the induction of Ag-specific unresponsiveness. Treatment of dendritic cell cultures with recombinant HSP27 caused in the up-regulation of IL-1beta, TNF-alpha, IL-6, IL-12p70, and IL-12p40 but not IL-23p19, which was inhibited when Abs to HSP27 were added. The 1-fluoro-2,4-dinitrobenzene-conjugated dendritic cells that had been treated with HSP27 had an increased capacity to initiate contact hypersensitivity responses compared with control dendritic cells. This augmented capacity required TLR4 signaling because neither cytokine production by dendritic cells nor the increased induction of contact hypersensitivity responses occurred in TLR4-deficient C3H/HeJ mice. Our findings indicate that a cascade of events occurs following initial interaction of hapten with the skin that includes increased activity of HSPs, their interaction with TLR4, and, in turn, increased production of cytokines that are known to enhance Ag presentation by T cells. The results suggest that HSPs form a link between adaptive and innate immunity during the early stages of contact hypersensitivity.
Article
Skin is one of the main targets for reactive oxygen species; thus, reactive oxygen species-induced damage and protein and lipid modifications occur, and skin can undergo a wide array of diseases, from photosensitivity to cancer. In this study, human dermal fibroblasts exposed to hydrogen peroxide (0-1000 micromol/L) exhibited a marked increase in both protein carbonyls and 4-hydroxy-2-nonenal, which are indices of protein and lipid oxidation, respectively. An amount of 25 micromol/L ferulic acid ethyl ester, a well-known nutritional antioxidant, significantly counteracted both protein and lipid oxidation and reduced the loss in cell viability elicited by 500 micromol/L of hydrogen peroxide. A common way for cells to react to oxidative stress is up-regulation of vitagenes. To the vitagene family belong the heat shock proteins heme oxygenase-1 and heat shock protein-70, which are involved in the cellular defense against oxidative stress by different mechanisms. The administration of 25 micromol/L ferulic acid ethyl ester significantly decreased hydrogen peroxide-induced protein and lipid oxidation. Dermal fibroblasts exposed to 25 micromol/L ferulic acid ethyl ester in the presence of 500 micromol/L hydrogen peroxide showed an increased level of both heme oxygenase-1 and heat shock protein-70 compared with dermal fibroblasts treated with hydrogen peroxide alone. These findings provide evidence for the protective role of vitagenes in free radical-induced skin damage and highlight the potential protective use of nutritional antioxidants, such as ferulic acid and its derivatives.
Article
Heat shock protein 72 (HSP72) is involved in the myocardial self-preservation system under several conditions such as ischemia-reperfusion injury or late preconditioning. However, its mechanism is not fully understood. Ecto-5'-nucleotidase is a key enzyme for synthesizing adenosine and plays an important role in ischemic preconditioning. In this study, we tested the hypothesis that ecto-5'-nucleotidase plays a role in the cardioprotection of HSP72. Rat hearts (H group, n=6) were transfected with HSP72 gene by an intracoronary infusion of hemagglutinating virus of Japan (HVJ)-liposome complex. Control hearts (C group, n=6) were transfected with the beta-galactosidase gene. Following 30 min of normothermic ischemia, grafts were reperfused using Langendorff apparatus. The activity of ecto-5'-nucleotidase was significantly higher in H group than C group both before and after ischemia-reperfusion (H vs. C; 0.51+/-0.05 vs. 0.29+/-0.06, and 1.41+/-0.15 vs. 0.85+/-0.11 nmol/mg protein/min, P<0.05). H group also showed significant better functional recoveries than C group (P<0.05), as well as less creatine phosphokinase leakage (4.4+/-2.8 vs. 14.2+/-3.4 mU/min, P<0.05) and higher adenosine release (247.5+/-35.1 vs. 54.3+/-1.7 pmol/min, P<0.05). Administration of alpha,beta-methylene adenosine diphosphate (AMP-CP), an inhibitor of ecto-5'-nucleotidase, significantly diminished the tolerance to ischemia-reperfusion injury in H group (P<0.05). These results demonstrated that ecto-5'-nucleotidase activated by an overexpression of HSP72 attenuated ischemia-reperfusion injury in the rat myocardium. They suggest that ecto-5'-nucleotidase plays a role in the cardioprotective effects of HSP72 in rat hearts.
Article
Background: Heat shock proteins (HSPs) are expressed by most living cells and play fundamental roles in many biological processes. Their synthesis increases by a variety of stresses in order to enable cellular survival. Although it is known that they play an important role in immune and inflammatory responses of the skin, the role of HSPs in the pathogenesis of skin diseases has been studied in only limited skin diseases. Lichen planus (LP) is a relatively common papulosquamous dermatosis, and cell-mediated immunity plays an important role in its pathogenesis. Although an altered expression of certain HSPs was reported in oral LP lesions, the expression of HSPs in cutaneous lesions of LP has not been investigated. In this immunohistochemical study, we aimed at investigating the role of HSPs in the pathogenesis of LP by studying whether there is any difference in HSP expression in cutaneous lesions of LP when compared to normal skin and psoriasis vulgaris (PV). Methods: Formalin-fixed paraffin-embedded skin biopsy specimen blocks from LP patients (n = 39), patients with psoriasis (n = 20), and normal skin controls (n = 20) were used in the study. Antibodies to HSPs 60 and 70 were applied immunohistochemically by using streptavidin-biotin-horseradish peroxidase complex. An immunoreactivity intensity distribution index (IRIDI) was calculated to express the proportion of the immunoreactive cells as well as the staining intensity in different layers of the epidermis. Results: The mean IRIDI scores for HSP60 expression in the basal, suprabasal, and superficial epidermal layers of cutaneous LP were moderately higher than those of normal skin, but not different from those of PV skin. These scores for HSP70 in lesions of LP were moderately lower than those for normal skin in the basal layer, but not significantly different from normal in the other two layers. Scores for HSP70 in PV lesions were markedly lower in all three layers. In the cells of the inflammatory infiltrates (mostly lymphocytes), HSP60 scores for LP were moderately higher, compared to those for PV, whereas scores for HSP70 were much lower for LP and very much lower for PV. Conclusions: Significantly altered levels of HSP proteins were found in cutaneous LP lesions in comparison with normal skin and psoriasis, suggesting the role of HSPs in the pathogenesis of LP.
Article
Proteomics provides a powerful approach for screening alterations in protein expression and post-translational modification associated with particular human diseases. In this study, the analysis of protein expression was focused on malignant melanoma in order to determine the candidate genes involved in tumour progression. The proteomes of cultured melanocytes and of cell lines from primary and metastatic lesions of one malignant melanoma patient were profiled using two-dimensional electrophoresis (2-DE) and mass spectrometry. Differentially expressed proteins were confirmed by 2-DE and mass spectrometry on an additional four malignant melanoma cell lines. Total RNA from the first subset of cell lines was used for quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) of the candidate genes identified after proteomics analysis. A very high similarity was observed in the 2-DE maps of two malignant melanoma cell lines derived from primary and secondary lesions of the same patient. Mass spectrometry identified 37 proteins which were found to be more abundant in tumour cells in comparison with control melanocytes (as confirmed on additional cell lines), with a relatively high prevalence of stress proteins. Eight candidate genes (PRDX2, HSP27, HSP60, HSPA8, HSP9B, STIP1, PDI and P4HB) were further characterized by evaluating their messenger RNA expression levels through real-time RT-PCR analysis. Overexpression of HSP27, HSP60 and HSPA8 and downregulation of PRDX2 were observed in cells from metastatic malignant melanoma in comparison with those from primary melanoma. Although further investigations with larger numbers of paired normal and tumour samples are needed, our findings strongly suggest that the dysregulation of stress pathways may be involved in melanoma progression.
Article
Skin wounds, especially chronic wounds such as pressure and diabetic ulcers, are posing a huge threat to healthcare systems worldwide. Thus, novel strategies to realize effective wound therapy are nowadays extremely needed. Herein, we are the first to report a combination therapy of electric stimulation and heat for wound healing just using a facile piezoelectric and photothermal dual functional film. The film, name as [email protected], is simply fabricated by coating polydopamine on a chitosan film. [email protected] generates electric voltages under mechanical pressure and shows photothermal effect with irradiation of near-infra red (NIR) light. In vivo experiment with rat full-thickness wound model validates that [email protected] with NIR can effectively promote wound regeneration due to the generation of electric stimulation and heat on wounds. The healing mechanism of [email protected] is further investigated, and the result demonstrates that [email protected] enhances wound healing through a pathway in which heat shock protein 90 (Hsp90) and hypoxia-inducible factor 1α (HIF-1α) participate. These results indicate that [email protected] is a promising wound dressing candidate for clinical application in the future.
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Heat shock proteins (HSPs), a large group of highly evolutionary conserved proteins, are considered to be main elements of the cellular proteoprotection system. HSPs are encoded by genes activated during the exposure of cells to proteotoxic factors, as well as by genes that are expressed constitutively under physiological conditions. HSPs, having properties of molecular chaperones, are involved in controlling/modulation of multiple cellular and physiological processes. In the presented review, we summarize the current knowledge on HSPs in the biology of epidermis, the outer skin layer composed of stratified squamous epithelium. This tissue has a vital barrier function preventing from dehydratation due to passive diffusion of water out of the skin, and protecting from infection and other environmental insults. We focused on HSPB1 (HSP27), HSPA1 (HSP70), HSPA2, and HSPC (HSP90), because only these HSPs have been studied in the context of physiology and pathophysiology of the epidermis. The analysis of literature data shows that HSPB1 plays a role in the regulation of final steps of keratinization; HSPA1 is involved in the cytoprotection, HSPA2 contributes to the early steps of keratinocyte differentiation, while HSPC is essential in the re-epithelialization process. Since HSPs have diverse functions in various types of somatic tissues, in spite of multiple investigations, open questions still remain about detailed roles of a particular HSP isoform in the biology of epidermal keratinocytes.
Article
Methomyl (MET) is a carbamate insecticide which is used as a substitute for organophosphorus compounds to protect crops against insects. The present study aims to evaluate the cytoprotection response of pigment cells and heat shock protein 70 (HSP70) after exposure to MET during the tadpole developmental stages of the Arabian toad, Bufo arabicus. Three developmental larval stages of the toad were selected and divided into two groups; Control and MET-exposed (MET-EX) tadpoles (10ppm). MET-EX tadpoles showed an increased number of pigment cells in the liver, kidney, anterior eye chamber, and skin tissues as compared to the control. The glycogen content in the developing liver and muscles (myotomes) of MET-EX tadpoles was decreased as compared to the control. In the MET-EX tadpoles, immunohistochemical staining showed an increase of HSP70 expression in the liver hepatocytes, the nucleated red blood cells (nRBC) in kidney glomeruli, the iridocorneal angle of anterior eye chamber, and the skin as compared to the control. The current study concluded that pigment cells and HSP70 represented a cytoprotecting response against MET insecticide during the organ development of B. arabicas tadpoles. Therefore, MET use should be regularly monitored in the environment to protect animals and human from exposure to this insecticide.
Article
Transforming growth factor‐β (TGF‐β) and heat shock protein 70 (HSP70) are important for the hair follicle (HF) cycle, but it is unclear whether they participate in HF regression in yak skin. In this study, we investigated the role of TGF‐β, TGF‐βRII, and HSP70 in the transition from anagen to catagen of HFs. The results showed that TGF‐β2 transcription was significantly higher than that of TGF‐β1 and TGF‐β3 in the same periods. Meanwhile, the expressions of TGF‐β2, TGF‐βRII, and caspase‐3 were higher in the catagen phase than that in mid‐anagen, and some TGF‐βRII‐positive HF cells were terminal deoxynucleotidyl transferase‐mediated deoxyuridine triphosphate‐biotin nick end labeling (TUNEL)‐positive. Moreover, the HSP70 protein levels in mid‐anagen were higher than those in late‐anagen and catagen. These results suggested that TGF‐β2 plays a major role in catagen induction in yak HFs, which might be achieved via TGF‐βRII‐mediated apoptosis in HF epithelial cells. In contrast, HSP70 might protect epithelial cells from apoptosis and ultimately inhibit HF regression. In conclusion, TGF‐β2 has positive effects, whereas HSP70 has negative effects, on catagen induction.
Article
2,3,7,8-tetrachlordibenzo-p-dioxin (TCDD) is an environmental pollutant that can cause various toxic effects, including chloracne, metabolic syndrome, and immune suppression. Most of the toxicity associated with TCDD is mediated through activation of the aryl hydrocarbon receptor (AHR). Recent research has suggested the presence of a wide-range of interindividual variability in TCDD-mediated suppression of the Immunoglobulin-M (IgM) response across the human population. In an attempt to identify putative modifiers of AHR-mediated immunosuppression beyond the AHR, B cells were isolated from a panel of genetically diverse mouse strain to scan for modulators that drive inter-strain differences in TCDD-mediated suppression of the IgM response. Results implicated a region of mouse Chromosome 1 near a gene encoding serine peptidase inhibitor, clade B, member 2 (Serpinb2) whose human ortholog is plasminogen activator inhibitor 2 (PAI2). Further downstream analyses indicated that Serpinb2 is dysregulated by TCDD and, furthermore, that B cells from Serpinb2-/- mice are significantly more sensitive to TCDD-mediated suppression as compared to littermate controls. This study suggests a protective role of Serpinb2 within TCDD-mediated immunosuppression and, furthermore, a novel function of Serpinb2-related activity in the IgM response.
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Human skin, eye and hair color rely on the production of melanin, depending on its quantity, quality, and distribution, Melanin plays a monumental role in protecting the skin against the harmful effect of ultraviolet radiation and oxidative stress from various environmental pollutants. However, an excessive production of melanin causes serious dermatological problems such as freckles, solar lentigo (age spots), melasma, as well as cancer. Hence, the regulation of melanin production is important for controlling the hyper-pigmentation. Melanogenesis, a biosynthetic pathway to produce melanin pigment in melanocyte, involves a series of intricate enzymatic and chemical catalyzed reactions. Several extrinsic factors include ultraviolet radiation and chemical drugs, and intrinsic factors include molecules secreted by surrounding keratinocytes or melanocytes, and fibroblasts, all of which regulate melanogenesis. This article reviews recent advances in the development of melanogenesis inhibitors that directly/indirectly target melanogenesis-related signaling pathways. Efforts have been made to provide a description of the mechanism of action of inhibitors on various melanogenesis signaling pathways.
Article
Introduction Burn injury is common and depth is one measure of severity. Although the depth of burn injury is determined by many factors, the relationship between the temperature of the injurious agent and exposure duration, known as the time-temperature relationship, is widely accepted as one of the cornerstones of burn research. Moritz and Henriques first proposed this relationship in 1947 and their seminal work has been cited extensively. However, over the years, readers have misinterpreted their findings and incorporated misleading information about the time-temperature relationship into a wide range of industrial standards, burn prevention literature and medicolegal opinion. Aim The purpose of this paper is to present a critical review of the evidence that relates temperature and time to cell death and the depth of burn injury. These concepts are used by researchers, burn prevention strategists, burn care teams and child protection professionals involved in ascertaining how the mechanism of burning relates to the injury pattern and whether the injury is consistent with the history. Review methods This review explores the robustness of the currently available evidence. The paper summarises the research from burn damage experimental work as well as bioheat transfer models and discusses the merits and limitations of these approaches. Review findings There is broad agreement between in vitro and in vivo studies for superficial burns. There is clear evidence that the perception of pain in adult human skin occurs just above 43 °C. When the basal layer of the epidermis reaches 44 °C, burn injury occurs. For superficial dermal burns, the rate of tissue damage increases logarithmically with a linear increase in temperature. Beyond 70 °C, rate of damage is so rapid that interpretation can be difficult. Depth of injury is also influenced by skin thickness, blood flow and cooling after injury. There is less clinical evidence for a time-temperature relationship for deep or subdermal burns. Bioheat transfer models are useful in research and becoming increasingly sophisticated but currently have limited practical use. Time-temperature relationships have not been established for burns in children’s skin, although standards for domestic hot water suggest that the maximum temperature should be revised downward by 3–4 °C to provide adequate burn protection for children. Conclusion Time-temperature relationships established for pain and superficial dermal burns in adult human skin have an extensive experimental modeling basis and reasonable clinical validation. However, time-temperature relationships for subdermal burns, full thickness burns and burn injury in children have limited clinical validation, being extrapolated from other data, and should be used with caution, particularly if presented during expert evidence.
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Objective This experiment was conducted to study the histological characteristics, age-related thickness changes, and expression of HSPs in the skin of yak. Methods A total of 20 yaks (10 males and 10 females) were used. Different regions of the normal skin of three different ages (newborn, half-year-old and adult) of yaks were harvested for histological study and thickness measurement. Biopsy samples were taken from the scapula regions of the skin from the same five approximately 1-year-old yaks during the hair cycle (telogen, anagen and catagen). RT-PCR, western blot and immunohistochemistry methods using the mRNA and protein levels were used to detect the expression of HSP27, HSP70 and HSP90. RT-PCR method was used to detect the mRNA expression of CGI-58 and KDF1. The IPP6.0 software was used to analyze the immunohistochemistry and measure the thickness of the skin. Results The general histological structure of hairy yak skin was similar to other domestic mammals. The unique features included prominent cutaneous vascular plexuses, underdeveloped sweat glands, a large number of nasolabial glands in the nasolabial plate, and hair follicle groups composed of one primary follicle and several secondary follicles. The skin, epidermis and dermis thickness did vary significantly between different body regions and different ages. The thickness of the skin, epidermis and dermis increased from newborn to adult in yaks. Yak skin thickness decreased from dorsally to ventrally on the trunk. The skin on the lateral surface was thicker than the skin on the medial surface on the limbs. HSP27, HSP70 and HSP90 showed different expression patterns during the hair cycle using RT-PCR, western blot and immunohistochemistry methods. The expression of HSP27 mRNA and protein in the anagen stage was the highest, followed by the catagen stage, and the expression in the telogen stage was the lowest. The expression of HSP70 mRNA and protein in the telogen stage was the highest, followed by the anagen stage, and the expression in the catagen stage was the lowest. The expression of HSP90 mRNA and protein in the anagen stage was the highest, followed by the telogen stage, and the expression in the catagen stage was the lowest. HSPs were mainly expressed in the outer root sheath of hair follicle during the hair cycle, also expressed in epidermis, sebaceous gland and sweat gland in the skin of Yak. The expression of CGI-58 mRNA in the anagen stage was the highest, followed by the catagen stage, and the expression in the telogen stage was the lowest. The expression of KDF1 mRNA in the telogen stage was the highest, followed by the catagen stage, and the expression in the anagen stage was the lowest. Meaning In this study, we examined and fully described the histology of normal skin in Yak and measured the skin thickness of different ages and different regions in Yak. These data may be useful to better understand and appreciate the adaptability features of yak skin. Our investigation reports the expression patterns of HSPs in yak skin for the first time. The different expression pattern of HSPs during the hair cycle suggests they may play different roles in yak hair follicle biology.
Article
Background: Plasmacytoid dendritic cells (pDC) are a subset of DC specialized in the production of type I interferon (IFN-α/β) and involved in various cutaneous inflammatory and autoimmune disorders, such as cutaneous lupus (CLE) and vitiligo. Heat-Shock proteins (HSP) are molecular chaperones essential for maintaining cellular functions, but can act as a danger signal during inflammation. Objectives: To decipher the role of HSP70 in the production of IFNα by pDC in CLE and vitiligo. Methods: Expression of HSP70 and CD123(+) pDC was analyzed by immunohistochemistry or immunofluorescence in CLE and vitiligo skin samples. Flow cytometry was performed to analyze expression of HSP70 receptors, activation markers on pDC and DNA uptake by pDC in the presence of HSP70. Impact of HSP70 on DNA-induced IFNα secretion by pDC was evaluated by enzyme-linked immunosorbent assay (ELISA). The effect of IFNα on CXCL9/10 gene and protein expression by keratinocytes was determined by real time PCR and ELISA. Results: Infiltration of pDC in CLE and progressive vitiligo was primarily located in the epidermis, close to keratinocytes expressing HSP70. In vitro experiments revealed that pDC expressing HSP70 receptor LOX-1 were able to aggregate HSP70. Exogenous HSP70 induced activation of pDC and increased the uptake of exogenous DNA. Furthermore, HSP70 potentiated DNA-induced IFNα production by pDC. Lastly, IFNα induced expression of CXCL9 and CXCL10 by keratinocytes. Conclusions: These data demonstrate that interaction between HSP70 and pDC in CLE and vitiligo is a prerequisite for the enhancement of IFNα production, and could be an interesting target. This article is protected by copyright. All rights reserved.
Article
Skin is most important environmental interface providing a protective envelope to animals. It's always under the influence of both internal and external stressors. Heat shock proteins (HSP) are highly conserved stress proteins which play crucial roles in environmental stress tolerance and thermal adaptation. Present study was planned to observe the relative mRNA expression of inducible (HSP70.1 and HSP70.2) and constitutive (HSP70.8) HSP in skin of zebu (Tharparkar) and crossbred (Karan Fries) cattle during different seasons. Skin biopsies were collected from rump region of each animal, aseptically during winter, spring and summer season. Quantitative real time polymerase chain reaction was performed to examine the gene expression of constitutive (HSP70.8) and inducible (HSP70.1 and HSP70.2) HSP in skin of both the breeds during different seasons. Present study observed higher expression of both constitutive and inducible HSP genes in both the breeds during summer and winter than spring season, but magnitude of increase was higher during summer than winter. During summer season, expression pattern of HSPs in skin showed breed differences, where constitutive HSP expression was higher in Tharparkar than Karan Fries and that of inducible HSP was higher in Karan Fries than Tharparkar. Hence, present study suggested that HSP may be conveniently used as biomarkers for assessing protective response of skin against heat stress in zebu and crossbred cattle. Variation in expression between breeds is associated with their heat tolerance and thermal adaptability. In summary, skin of zebu cattle (Tharparkar) is more resistant to summer stress than crossbred (Karan Fries), providing greater protection against heat stress during summer season. Superior skin protective mechanism of zebu (Tharparkar) than crossbred (Karan-Fries) cattle against heat stress may contribute to superior adaptability of zebu cattle to tropical climatic conditions than crossbreed.
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The heat shock or stress response of Escherichia coli has evolved in order to detect and deal with the presence of unfolded, misfolded, damaged or aggregated polypeptide chains. At the present, two major regulons are known to control this response. The “classical” heat shock regulon has evolved to deal with intracellular protein perturbations and is under the positive control of the σ32 transcription factor (the rpoH gene product) and the negative control of some of the heat shock proteins themselves. The newly discovered second heat shock regulon has evolved to deal with protein misfolding/aggregation/imbalance in the outer cellular compartments. It is under the positive control of the σE transcription factor (the rpoE gene product). The two heat shock regulons appear to be interconnected, inasmuch as the σE factor participates in the transcriptional regulation of the σ32-encoding gene, especially at very high temperatures.
Article
Toll-like receptors (TLRs) mediate not only innate immunity against infection and but also sterile inflammation triggered by endogenous molecules. We conducted a comparative study of the different inflammatory responses induced by repetitive ultraviolet (UV) B irradiation in wild-type (WT) and TLR2 knock-out (KO) mice, to provide in vivo evidence of the role of TLRs in mediating UVB-induced responses. UVB-induced inflammatory responses were less severe in TLR2 KO mice than in WT mice after 6 weeks of repeated UVB irradiation. UVB-treated TLR2 KO mice displayed less prominent erythema and scaling, and histopathology showed significantly thinner skin and less inflammatory cell infiltration than that in WT mice. UVB-induced expression of heat shock protein 70 (an endogenous ligand of TLR2) was lower in TLR2 KO mice. Quantitative RT-PCR revealed significantly lower gene expression levels of UVB-induced interleukin (IL)-1β, IL-6, and matrix metalloproteinase (MMP)-13 in TLR2 KO mice. TLR2 KO mice also showed significantly lower protein level expression of UVB-induced IL-1β in ELISA and MMP-13 in western blots. Our study demonstrated that TLR2 was associated with inflammatory responses to repetitive UVB irradiation in C57/BL6 mice. Moreover, it suggests that the role of TLR2 in the cutaneous response of UV irradiation and in developing new agents for modulating the effects of UV irradiation should be considered.This article is protected by copyright. All rights reserved.
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Very high frequency ultrasound (VHF-US) is new therapy method with a broad application spectrum in dermatology and aesthetic medicine. In this method, ultrasound waves with frequencies over 10 MHz, which were for a long time only used in ultrasound diagnostics, are applied for therapeutic purposes. Such US waves demonstrate specific biophysical efficiencies which warrant their application for the treatment of the skin efflorescences, chronic wounds and hypertrophic scars as well as in anti-aging and skin improvement procedures in aesthetic medicine. VHF-US can be applied not only for stand-alone treatments, but also as a supportive pre- and posttreatment method in combination with laser, radiofrequency currents, injection lipolysis, etc. as well as in aesthetic plastic surgery.
Article
Background Atopic dermatitis is a chronic, relapsing inflammatory disease of the skin. Current therapy is not curative and recalcitrant disease is a big stress and challenge for parents and physicians. This study explored the potential role of Heat shock protein 70 (HSP-70) and its anti-inflammatory effects on keratinocyte under TH2 environment.Methods Human keratinocyte cell line (HaCa T) was stimulated with IL-4, IL-13, and TNF-α to synthesize and secrete thymic stromal lymphopoietin (TSLP), an important cytokine of immuno-pathogenesis in atopic dermatitis. Heat shock was performed by immersing the cell-contained flash into a water bath of 45°C for 20 minutes. Cell viability, TSLP expression and secretion of HaCa T cells were measured and compared. Possible regulatory mechanisms influencing the expression of TSLP, such as the STAT6 and NF-κB signal pathways, were investigated.ResultsHeat shock treatment induced intracellular HSP-70 expression in HaCa T cells without affecting cell viability. The induced expression and secretion of TSLP in HaCa T cells were suppressed by heat shock. The NF-κB signal pathway was inhibited by heat shock, leading to decreased TSLP expression and secretion.Conclusion Heat stress induced HSPs can significantly reduce the production and secretion of TSLP from HaCaT cells under Th2 environment. Thus, the evidence highlights the potential role of HSP-70 for atopic dermatitis in the future.This article is protected by copyright. All rights reserved.
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Nowadays, people are living much longer than they used to do, however they are not free of disabilities and diseases, which still represent the dark side of aging and longevity. Aging is a relentless process that affects all cells, tissues, organs, and organisms, diminishing homeostasis and increasing organism vulnerability [1].
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The pathogenesis of canine atopic dermatitis (AD) involves dysfunction of the adaptive immune system. Recent evidence suggests that nonantigen-specific inflammatory elements may play a role in the development and perpetuation of canine AD. The objective of this review is to provide an update on recent advances in the understanding of the role of innate immune cells, keratinocytes, lipid metabolism and nutrition in the pathogenesis of AD in dogs. Citation databases, abstracts and proceedings from international meetings published between 2001 and 2013 are reviewed in this update. Where necessary, older articles are included for background information. Members of the innate immune system (including dendritic cells, Langerhans cells and mast cells) and keratinocytes interact with each other and with environmental antigens during both induction and effector phases of atopic inflammation. The responses of these cells and associated noncellular factors (such as complement and protease-activated receptors) to environmental stimuli influence the entire future course of the immune response to a given agent. Abnormalities in lipid metabolism may also influence the pathogenesis of canine AD via the production of inflammatory mediators and by alteration of epidermal barrier function and antigen presentation. However, a lack of fully controlled studies precludes definitive interpretation of these data. Evidence indicates that the cells and noncellular components of the innate immune system and the epidermis may play critical roles during both the sensitization and the effector phases of canine AD. Derangements in lipid metabolism may be involved in the pathogenesis of AD in dogs, but additional controlled studies are required in this area. © 2015 ESVD and ACVD.
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The expression of heat shock proteins (HSPs), particularly HSP70, is receiving considerable attention in the field of cosmetics, particularly given our recent report that ultraviolet-induced melanin production, skin damage and wrinkle formation were all suppressed in transgenic mice expressing HSP70. In the present study, we searched for HSP70-inducers from a library of herbal extracts that have already been approved as quasi-pharmaceutical products in Japan. We selected an ethanol extract of Arnica montana (A. montana), based on its high HSP70-inducing activity and low cytotoxicity. Cell viability was determined by MTT method and expression of HPS70 was monitored by immunoblotting analysis. From the extract, we purified and identified eight sesquiterpene lactones (AM1-8) as HSP70-inducers, among which AM-2 (helenalin 2-methylbutyrate) was selected due to its good HSP70-inducing properties and low cytotoxicity. Treatment of cultured mouse melanoma cells with AM-2 or A. montana extract up-regulated the expression of HSP70 in a dose-dependent manner. This treatment also activated heat shock factor-1, a transcription factor for hsp genes. Furthermore, pre-treatment of cells with AM-2 or A. montana extract decreased melanin production and expression and activity of tyrosinase. These results suggest that AM-2 and A. montana extract could be beneficial for use in hypopigmenting cosmetics as a consequence of their stimulatory effects on HSP70 expression. Copyright © 2015 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.
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Designing biomaterials capable of biomimicking wound healing and skin regeneration has been receiving increasing attention recently. Some biopolymers behave similarly to the extracellular matrix (ECM), supporting biointerfacial adhesion and intrinsic cellular interactions. Polydopamine (PDA) is a natural bioadhesive and bioactive polymer that endows high chemical versatility, making it an exciting candidate for a wide range of biomedical applications. Moreover, biomaterials based on PDA and its derivatives have near-infrared (NIR) absorption, excellent biocompatibility, intrinsic antioxidative activity, antibacterial activity, and cell affinity. PDA can regulate cell behavior by controlling signal transduction pathways. It governs the focal adhesion behavior of cells at the biomaterials interface. These features make melanin-like PDA a fascinating biomaterial for wound healing and skin regeneration. This paper overviews PDA-based biomaterials' synthesis, properties, and interactions with biological entities. Furthermore, the utilization of PDA nano- and microstructures as a constituent of wound-dressing formulations is highlighted.
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Among raised dermal scar types, keloid (KS) and hypertrophic scars (HS) are considered to present clinical similarities, but there are no known specific biomarkers that allow both scar types to be easily distinguished. Development and progression of raised dermal scars comprises the activation of several molecular pathways and cell defence mechanisms leading to elevated extracellular matrix component synthesis, delayed apoptosis, altered migration and differentiation. Therefore, the aim here was to identify biomarkers that may differentiate between KS and HS compared to normal skin (NS). To achieve this aim, NS (n = 14), KS (n = 14) and HS (n = 14) biopsies were evaluated using histology by H&E staining. Tissue biopsies and primary fibroblasts (passages 0-4) were employed to assess the gene expression levels of 21 biomarkers selected from our previous microarray studies using qRT-PCR. Finally, protein expression was evaluated using In-Cell Western Blotting in primary fibroblasts (p 0-4). Our results demonstrated that out of the 21 biomarkers screened at mRNA and protein levels, α2β1-integrin, Hsp27, PAI-2, MMP-19 and CGRP showed significantly higher expression (p < 0.05) in KS compared to NS and HS. Additionally, these five key biomarkers were found to be significantly higher (p < 0.05) at mRNA level in KS taken from the sternum, a region known to be subjected to high mechanical forces in the body during the performance of daily movements. In conclusion, our findings offer potential molecular targets in raised dermal scars differentiation. Future targeted research may allow provision of diagnostic and prognostic markers in keloid versus hypertrophic scars
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Although a wide variety of protein profiles have been extensively constructed via proteomic analysis, the comprehensive proteomic profiling of the skin, which is considered to be the largest organ of the human body, is still far from complete. Our efforts to establish the functional skin proteome, a protein database describing the protein networks that underlie biological processes, has set in motion the identification and characterization of proteins expressed in the epidermis and dermis of the BALB/c mice. In this review, we will highlight various cutaneous proteins we have characterized and discuss their biological functions associated with skin distress, immunity, and cancer. This type of research into functional skin proteomics will provide a critical step toward understanding disease and developing successful therapeutic strategies.
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Heat stress and ultraviolet light in the UVB range ("sunburn spectrum," 290-320 nm) were found to alter the synthesis of specific proteins in cultured keratinocytes derived from mouse skin. Using giant two-dimensional gels, approximately 2,000 cellular polypeptides labeled with [35S]methionine at 4-5 h after exposure to heat or to UVB were analyzed. Cells conditioned at sublethal temperatures (42 degrees C for 1 h, or 47 degrees C for 15 min) developed thermotolerance, while cells conditioned with UVB did not develop thermotolerance. Under these heat or UVB conditions, 19 stress proteins were observed. Proteins fell into three classes based upon their inducibility by heat or UVB, dose-response, and induction mechanism (transcriptional versus post-transcriptional) as defined by metabolic blockade with cordycepin (3'-deoxyadenosine). Class 1 proteins were inducible only by heat shock. They included three major heat-shock proteins (hsp 72, hsp 78, hsp 90) and a 42.5-kDa, pI 5.43 protein, and all were induced at the transcriptional level. Class 2 proteins were inducible by heat and by UVB. These included hsp 110 and eight additional polypeptides. All but one were affected by heat at the post-transcriptional level and were induced by UVB at both low (20 mJ/cm2) and high (80 mJ/cm2) doses. Class 3 proteins were inducible only at high UVB doses (survival < 10%). Class 1 and Class 2 proteins could be functionally involved in thermotolerance, while Class 3 proteins are more likely related to damage or cell death.
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Myocardial protection and changes in gene expression follow whole body heat stress. Circumstantial evidence suggests that an inducible 70-kD heat shock protein (hsp70i), increased markedly by whole body heat stress, contributes to the protection. Transgenic mouse lines were constructed with a cytomegalovirus enhancer and beta-actin promoter driving rat hsp70i expression in heterozygote animals. Unstressed, transgene positive mice expressed higher levels of myocardial hsp70i than transgene negative mice after whole body heat stress. This high level of expression occurred without apparent detrimental effect. The hearts harvested from transgene positive mice and transgene negative littermates were Langendorff perfused and subjected to 20 min of warm (37 degrees C) zero-flow ischemia and up to 120 min of reflow while contractile recovery and creatine kinase efflux were measured. Myocardial infarction was demarcated by triphenyltetrazolium. In transgene positive compared with transgene negative hearts, the zone of infarction was reduced by 40%, contractile function at 30 min of reflow was doubled, and efflux of creatine kinase was reduced by approximately 50%. Our findings suggest for the first time that increased myocardial hsp70i expression results in protection of the heart against ischemic injury and that the antiischemic properties of hsp70i have possible therapeutic relevance.
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To elucidate cellular concepts for protection against ultraviolet (UV) light we investigated the effect of heat shock protein 70 (hsp70) overexpression on cell viability and on the secretion of UV-inducible immunological cytokines. Transfected murine fibrosarcoma cells (WEHI-S), overexpressing hsp70 or a sham transfected control were used. Overexpression of hsp70 was sufficient to markedly increase cell viability upon treatment with UVB (290-320 nm). Since long wave UV (UVA, 320-400 nm) as well as UVB turned out to stimulate the release of O2- radicals we studied the cell viability upon oxidative stress. Hsp70 overexpression increased viability upon treatment with hydrogen peroxide or menadione, but had no influence on UV-induced O2- release. UV-light is known to upregulate immunologic and proinflammatory cytokines such as IL-1 and IL-6. Oxidative stress appeared to exert a similar effect. Hsp70 overexpression markedly decreased the release of IL-6 induced by UVA, UVB and oxidative stress. To test whether the hsp70 mediated suppression is confined to events caused by UV-light we determined IL-1-mediated effects. IL-1-induced IL-6 release was reduced by hsp70 overexpression, whereas the IL-1 mediated activation of nuclear factor kappa B was not affected. Our data suggests that hsp70 plays a central role not only in cell protection against UV-light, but also in the regulation of proinflammatory cytokine release induced by UV-exposure.
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Preservation of the chemical architecture of a cell or of an organism under changing and perhaps stressful conditions is termed homeostasis. An integral feature of homeostasis is the rapid expression of genes whose products are specifically dedicated to protect cellular functions against stress. One of the best known mechanisms protecting cells from various stresses is the heat-shock response which results in the induction of the synthesis of heat-shock proteins (HSPs or stress proteins). A large body of information supports that stress proteins--many of them molecular chaperones--are crucial for the maintenance of cell integrity during normal growth as well as during pathophysiological conditions, and thus can be considered "homeostatic proteins." Recently emphasis is being placed on the potential use of these proteins in preventing and/or treating diseases. Therefore, it would be of great therapeutic benefit to discover compounds that are clinically safe yet able to induce the accumulation of HSPs in patients with chronic disorders such as diabetes mellitus, heart disease or kidney failure. Here we show that a novel cytoprotective hydroxylamine derivative, [2-hydroxy-3-(1-piperidinyl) propoxy]-3-pyridinecarboximidoil-chloride maleate, Bimoclomol, facilitates the formation of chaperone molecules in eukaryotic cells by inducing or amplifying expression of heat-shock genes. The cytoprotective effects observed under several experimental conditions, including a murine model of ischemia and wound healing in the diabetic rat, are likely mediated by the coordinate expression of all major HSPs. This nontoxic drug, which is under Phase II clinical trials, has enormous potential therapeutic applications.
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Wound healing conditions generate a stressful environment for the cells involved in the regeneration process and are therefore postulated to influence the expression of heat shock proteins (Hsps). We have examined the expression of four Hsps (Hsp27, Hsp60, Hsp70 and Hsp90) and a keratin (keratin 6) by immunohistochemistry during cutaneous wound repair from Day 1 to Day 21 after wounding in the mouse. Hsps were constitutively expressed in normal mouse epidermis and their patterns of expression were modified during the healing process. The changes were not directly linked to the time course of the healing process but rather were dependent on the location of cells in the regenerating epidermis. In the thickened epidermis, Hsp60 was induced in basal and low suprabasal cells, Hsp70 showed a reduced expression, and Hsp90 and Hsp27 preserved a suprabasal pattern with an induction in basal and low suprabasal cells. All Hsps had a uniform pattern of expression in the migrating epithelial tongue. These observations suggest that the expression of Hsps in the neoepidermis is related to the proliferation, the migration, and the differentiation states of keratinocytes within the wound.
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Glucocorticoid receptors must be complexed with Hsp90 in order to bind steroids, and it has been reported that at least three other proteins, Hop, Hsc70, and a J-domain protein (either Hsp40 or Ydj1), are required for formation of active Hsp90-steroid receptor complex. In the present study, we reinvestigated activation of stripped steroid receptors isolated from either L cells or WCL2 cells. Surprisingly, we found, using highly purified proteins, that only Hsp90 and Hsc70 are required for the activation of glucocorticoid receptors in the presence of steroids; in the absence of steroids, either p23 or molybdate are also required as reported previously. Addition of Hop or Ydj1 had no affect on the rate or magnitude of the activation of the stripped receptors, and quantitative Western blots confirmed that neither Hop or Hsp40 were present in our protein preparations or in the stripped receptors. Furthermore, a truncated recombinant Hsp70 that does not bind Hop or Hsp40 was as effective as wild-type Hsp70 in activating stripped receptor. Since Hsc70 does not bind directly to Hsp90 but both proteins bind to Hop, it has been suggested that Hop acts as a bridge between Hsp90 and Hsp70. However, we found that after Hsc70 or Hsp90 bind directly to the stripped receptors, they are fully reactivated by Hsp90 or Hsc70, respectively. We, therefore, conclude that Hsp90 and Hsc70 bind independently to stripped glucocorticoid receptors and alone are sufficient to activate them to bind steroids.
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Members of the heat shock protein 70 (HSP70) family display a broad cellular localization and thus bind a repertoire of chaperoned peptides potentially derived from proteins of different cellular compartments. In this report, we show that HSP70 purified from human melanoma can activate T cells recognizing melanoma differentiation antigens in an antigen- and HLA class I-dependent fashion. HLA class I-restricted anti-melanoma T cells were susceptible to MHC-restricted, HSP70-dependent stimulation, indicating that HSP70 complexed peptides were able to gain access to the class I HLA presentation pathway. In addition, MHC matching between the melanoma cells used as a source of HSP and the responding T cells were not required, indicating that HSP70 activation may occur across MHC barriers. Besides the MHC-restricted and peptide-dependent activation pathway, HSP70 with no endogenous complexed peptides or HSP70 purified from antigen-negative cells was also able to induce IFN-gamma release by antimelanoma T cells by a MHC-independent mechanism. In this case, however, higher doses of HSP70 were required. The capacity to activate class I-restricted, antitumor T cells as well as antigen-presenting cells, together with the finding that the HSP70 chaperoned peptide repertoire includes melanoma-shared epitopes, holds promise for a HSP70-based cancer vaccine.
Chapter
In view of the fundamental role of stress proteins in the maintenance of protein homeostasis, it seems likely that malfunctions associated with members of stress protein families would have pathological effects. Such effects might be minimal under normal physiological conditions, but could be exacerbated at times when other disease stimuli trigger the requirement for local alterations in stress protein function in particular afflicted cells or tissues. During infection, it can be anticipated that the requirement of stress proteins for cell viability will be equally essential both for the pathogen and for the infected host. Just as stress proteins are essential in “normal” as well as stressed cells, it is clear that changes in stress protein expression will be associated with physiologically normal events accompanying infection as well as with any subsequent pathological events. In addition to the direct role of stress proteins in cell physiology, their potential medical influence is compounded by their ability to act as potent immunogens. Responses to microbial stress proteins are a prominent feature of the immune repertoire in patients and in experimental animals, and there has been wide discussion of the possibility that recognition of conserved, self-like, epitopes on such antigens could influence infectious and other diseases. Three broad hypotheses have been put forward concerning the relevance of immunological reactivity to stress proteins:
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This study aimed to evaluate a 815 nm diode-laser system to assist wound closure. It was proposed to determine an optimal fluence being able to accelerate and improve heating process without thermal damage after laser irradiation. Male hairless rats with dorsal skin incisions were used for the study. Different fluences were screened (76 to 346 J/cm2) in a first phase with clinical examination at 3, 7, 15 and 21 days after surgery. Best results were obtained for a fluence of 145 J/cm2 and 3 sec time of exposure. A second phase was conducted to valid these parameters with histological study and determination of tensile strength at 3, 7, 15 and 21 days after surgery. LASC was 4 times faster to process than conventional suture. In the laser group with an optimal fluence of 145 J/cm2, healing was accelerated. The resulting scar was more indiscernible than in the control groups. Histological aspect was better with continuous epidermis and dermis at 3 days in most cases. Tensile strength was 30 to 58% greater than in control groups (1141 g/cm2 at 7 days in the laser group versus 856 g/cm2 and 724 g/cm2 in the control groups, p < 0.001).
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Death receptors are a growing family of transmembrane proteins that can detect the presence of specific extracellular death signals and rapidly trigger cellular destruction by apoptosis. Expression and signaling by death receptors and their respective ligands is a tightly regulated process essential for key physiologic functions in a variety of organs, including the skin. Several death receptors and ligands, Fas and Fas ligand being the most important to date, are expressed in the skin and have proven to be essential in contributing to its functional integrity. Recent evidence has shown that Fas-induced keratinocyte apoptosis in response to ultraviolet light, prevents the accumulation of pro-carcinogenic p53 mutations by deleting ultraviolet-mutated keratinocytes. Furthermore, there is strong evidence that dysregulation of Fas expression and/or signaling contributes to the pathogenesis of toxic epidermal necrolysis, acute cutaneous graft versus host disease, contact hypersensitivity and melanoma metastasis. With these new developments, strategies for modulating the function of death receptor signaling pathways have emerged and provided novel therapeutic possibilities. Specific blockade of Fas, for example with intravenous immunoglobulin preparations that contain specific anti-Fas antibodies, has shown great promise in the treatment of toxic epidermal necrolysis and may also be useful in the treatment acute graft versus host disease. Likewise, induction of death signaling by ultraviolet light can lead to hapten-specific tolerance, and gene transfer of Fas ligand to dendritic cells can be used to induce antigen specific tolerance by deleting antigen-specific T cells. Further developments in this field may have important clinical implications in cutaneous disease.Keywords: apoptosis, death receptors, Fas, skin
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Si notato che shocks di temperatura possono indurre una variazione di puffing pattern in ghiandole salivari di Drosophila. Tali puffing sono perfettamente reversibili e rappresentano zone di intensa sintesi di RNA. Si notato che DNP e Na salicilato portano a simili variazioni di puffing pattern.
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Small stress proteins (sHsp) are molecular chaperones whose expression was shown to enhance the survival of mammalian cells exposed to numerous types of injuries that lead to death, including heat shock, oxidative stress as well as treatments with anti-cancerous and apoptosis-inducing agents. Here, a summary of the most recent results concerning the protective activity of this family of proteins against programmed cell death is presented. (1) sHsp enhance the survival of cells exposed to oxidative stress, a phenomenon which is linked to the ability of these proteins to decrease the intracellular level of reactive oxygen species in a glutathione dependent way. (2) sHsp protect against apoptosis mediated by different agents including staurosporine, etoposide and the Fas ligand. (3) An interesting and particular aspect of sHsp concerns their transient expression during the cell division to differentiation transition. In this context, sHsp expression was shown to be essential for preventing differentiating cells from undergoing apoptosis. Small stress proteins appear therefore as novel regulators that interfere with programmed cell death induced by different pathways.
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We have shown previously that human epidermal keratinocytes in situ and in vitro constitutively express high levels of the 72-kD heat shock protein (hsp72) and that hsp72 expression in these cells can be further induced with heat treatment. In the present study, we continue our investigation of the ultraviolet (UV) B protective effect of hyperthermic treatment and ask whether hsp72 is a mediator of heat-shock-induced UVB resistance. The results of our experiments demonstrate that heat treatment (42 degrees C for 4 h) before UVB exposure is able to increase significantly the UVB resistance of the epidermal carcinoma cell line A431. Heat-induced UVB resistance was most pronounced if the cells were exposed to UVB immediately after heat treatment. The protective effect was not detectable beyond a recovery period of 12 h. To investigate the role of hsp72 in hyperthermia-induced UVB resistance, we inhibited the expression of this protein using either a specific antisense oligodeoxynucleotide or quercetin, a flavonoid that has been shown to down-regulate hsp expression. Treatment with the oligomer as well as with quercetin significantly increased the susceptibility of A431 to UVB-induced damage and nullified the protective effect of heat preconditioning. A noncomplementary control oligodeoxynucleotide had no significant effect. These results indicate that heat treatment is able to induce a state of increased resistance to the deleterious effects of UVB in human keratinocytes in vitro. hsp72 is a molecular mediator of this protective effect, and its constitutive expression in human epidermal keratinocytes may be an important mechanism for the protection of human epidermis from UVB-induced damage.
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Recent advances in the biology of heat-shock proteins (hsps) are reviewed. These abundant and evolutionarily highly conserved proteins (also called stress proteins) act as molecular escorts. Hsps bind to other cellular proteins, help them to fold into their correct secondary structures, and prevent misfolding and aggregation during stress. Cytoplasmic hsp70 and hsp60 participate in complicated protein-folding pathways during the synthesis of new polypeptides. Close relatives of hsp70 and hsp60 assist in the transport and assembly of proteins inside intracellular organelles. Hsp90 may have a unique role, binding to the glucocorticoid receptor in a manner essential for proper steroid hormone action. Hsps may also be essential for thermotolerance and for prevention and repair of damage caused by ultraviolet B light. A unique class of T lymphocytes, the gamma delta T cells, exhibits a restricted specificity against hsps. These T cells may constitute a general, nonspecific immune mechanism directed against the hsps within invading organisms or against very similar hsps within invading organisms or against very similar hsps expressed by infected (stressed) keratinocytes. Immunologic cross-reactivity between hsps of foreign organisms and of the host may play a role in some autoimmune diseases. Although hsps are expressed in the skin, many questions remain about their role during injury, infection, and other types of cutaneous pathophysiology.
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Two components of sunlight, ultraviolet (UV) B (290-320 nm) and infrared (greater than 700 nm), each cause damage to the skin. However, we recently identified a protective response in which heat reduces UVB-induced killing of cultured keratinocytes. Here, this investigation is extended to the living epidermis. The effects of hyperthermic preconditioning upon UVB-induced apoptosis were studied morphologically with hematoxylin and eosin staining, and biochemically with TUNEL (terminal deoxynucleotide transferase nick-end labeling) to measure endonucleolytic cleavage of DNA in situ. Anesthetized SKH-1 hairless mice were exposed to UVB light (0 to 120 mJ/cm2), after which their skin was biopsied at 24 h and paraffin sections were stained with hematoxylin and eosin or with TUNEL. Apoptotic keratinocytes were found to increase after UVB in a dose-related manner. In contrast, if one flank of the mouse was pretreated at 40 degrees C for 1 h and both flanks subsequently were UVB-irradiated at 6 h, the resulting formation of apoptotic cells was reduced twofold or more in the heated flank. Protection appeared by 3 h, reached a maximum at 6 h, and disappeared by 12 h. In summary, heat induces a transient protective effect that reduces UVB-mediated death of keratinocytes in skin at physiologically attainable doses of heat and UVB.
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A field of research that began with a curious observation in Drosophila has resulted in a new understanding of how cells respond to sudden and adverse changes in their environment. In addition through the study of the structure/function of the stress proteins, especially those which function as molecular chaperones, new insights into the details by which proteins are synthesized and acquire their final biologically active conformation have been realized. Equally exciting is the progress being made as it relates the potential diagnostic and therapeutic applications of the stress-response proteins. The use of stress proteins as the next generation of vaccines and/or their use as potentially powerful adjuvants, capable of stimulating both T and B cell responses to a particular antigen of interest appear close to becoming a reality. One wonders how many more surprises are in store for us as we continue to explore this evolutionally conserved cellular stress response.
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Exposure of cells to elevated temperatures induces a physiologic response characterized by the synthesis of a specific set of proteins (heat shock or stress proteins, HSPs) mediating repair mechanisms and protection from cellular damage. In the present study upon immunohistochemistry using a specific monoclonal antibody, the constitutive and heat-induced expression of the 72-kD HSP (HSP72) in normal human skin and in human epidermal cell lines (KB, A431) was investigated. Normal (unstressed) epidermis and adnexal structures of normal human skin were found to constitutively express HSP72. In contrast, a substantial HSP72 expression could not be observed in the dermal cellular compartment. In vitro heat treatment of punch biopsies from normal skin (42 degrees C, 4 h) resulted in a further increase of epidermal HSP72 expression. In addition, dermal cells were found to be induced to express HSP72. To further evaluate the spontaneous HSP72 expression of epidermal cells two epidermoid carcinoma cell lines (A431, KB) were investigated. Upon immunohistochemistry and Western blot analysis a significant HSP72 expression could be detected in unstressed KB and A431 cells. In contrast, a human fibrosarcoma cell line (HT1080) was negative for HSP72 at 37 degrees C but upon heat treatment a strong induction was observed. Furthermore, Northern blot analysis using a cDNA probe specific for human HSP72 revealed a constitutive expression of HSP72 mRNA in both epidermal cell lines. These findings demonstrate a significant expression of the stress-inducible HSP72 in unstressed human skin as well as in epidermal cell lines, suggesting that HSP72 may inherently be involved in the protective function of normal human skin.
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Environmental exposure to UVB (290-320 nm) wavelengths of the solar spectrum causes major damage, including carcinogenesis, in the skin. Therefore, cellular responses that protect against UVB damage are of particular interest in cutaneous epithelial cells. In cultured keratinocytes, mild hyperthermia generates a classical stress response with acquired thermotolerance and elevated stress protein synthesis (E. V. Maytin, J. Biol. Chem., 267: 23189-23196, 1992). To test the ability of this stress response to protect against UVB damage, monolayers of primary murine keratinocytes or BALB/MK keratinocytes were heated at 42 degrees C for 1 h and then exposed to UVB at 6 h (typical dose, 40 mJ/cm2). Survival was assessed by fluorescein diacetate/ethidium bromide vital dye uptake and video microscopy. With heat-conditioning prior to UVB, a significant increase in both the percentage viability (2- to 3-fold) and in the absolute number of living (fluorescein diacetate-positive) cells was measurable at 24-48 h. Steady-state incorporation into [3H]DNA and 35S-protein, while suppressed immediately after UVB, showed greater recovery in heat-conditioned cultures compared to sham-conditioned cultures at 48 h. Increased metabolic activity was accompanied by increased proliferative potential since colonies of BALB/MK cells observed at 72 h were larger, more numerous, and more active in the uptake of 5-bromo-2'-deoxyuridine in heat-conditioned cultures. A time course for the development of UVB resistance showed maximal protection when heat and UVB were spaced approximately 6 h apart. Hyperthermic conditioning could induce UVB protection in nonproliferating cells, indicating that cell cycle arrest was not primarily responsible for the UVB-protective effect. In summary, hyperthermia induces a mechanism in epithelial cells which can ameliorate damage from UVB.
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Heat-shock proteins (HSPs), or so-called 'stress proteins' may play an important role in cutaneous pathophysiology. HSPs are a group of highly conserved molecules that are expressed by all cells when subjected to heat or other forms of physical or chemical stress. The physiological roles of stress proteins are varied and are important in stress and nonstress conditions. They bind to other cellular proteins and participate in protein folding pathways during stress and also during the synthesis of new polypeptides. HSPs are also essential for thermotolerance and for prevention and repair of damage caused in DNA after ultraviolet exposure. Although HSPs are expressed in the skin in both epidermis and dermis, HSPs may influence many other cellular processes in the inflammatory and immune skin response. Many authors have speculated on a link between HSPs and human skin disease characterized by inflammation and proliferation.
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Heat shock protein (hsp) induction by stressful stimuli such as heat and ischemia is known to protect cardiac cells from severe stress. The ability to induce hsp's in the heart directly by "nonstressful" means would potentially have important clinical implications. In noncardiac cells, the tyrosine kinase inhibitor herbimycin-A has been shown to induce the 72-kD hsp. We therefore examined whether herbimycin-A and another tyrosine kinase inhibitor, genistein, could induce 70-kD hsp's in primary cultures of rat neonatal cardiomyocytes, and whether these treatments protect against severe stress. Primary cardiomyocytes were incubated with herbimycin-A or genistein. hsp induction was measured 16-20 h later by Western blotting. Cell survival after subsequent lethal heat stress or simulated ischemia was assessed using trypan blue exclusion and released lactate dehydrogenase activity. Our results indicate that, in cardiac cells, herbimycin-A induces 70-kD hsp's but not hsp90, -60, -25, or glucose-regulated protein 78, whereas genistein has no effect on hsp's. Moreover, hsp induction correlated with the ability of herbimycin-A to protect cells against severe stress, whereas genistein has no protective effects. This suggests that herbimycin-A may induce 70-kD hsp's via a tyrosine kinase-independent mechanism. These results indicate the possibility of a pharmacological approach to HSP70 induction and cardiac protection, which may ultimately be of clinical relevance.
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Immunotherapy of mice with preexisting cancers with heat shock protein preparations derived from autologous cancer resulted in retarded progression of the primary cancer, a reduced metastatic load, and prolongation of life-span. Treatment with heat shock protein preparations derived from cancers other than the autologous cancer did not provide significant protection. Spontaneous cancers (lung cancer and melanoma), chemically induced cancers (fibrosarcoma and colon carcinoma), and an ultraviolet radiation-induced spindle cell carcinoma were tested, and the results support the efficacy of autologous cancer-derived heat shock protein-peptide complexes in immunotherapy of cancers without the need to identify specific tumor antigenic epitopes.
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In response to ultraviolet radiation (UVR), skin keratinocytes increase expression of heat shock proteins that can protect cells from stress-induced damage. This heat shock response is known to be transcriptionally regulated in eukaryotic cells exposed to certain forms of environmental stress. In the skin, absorption of ultraviolet B light occurs primarily in the epidermis, and therefore, using primary cultures of normal human epidermal keratinocytes, we have examined whether transcriptional activation of the hsp72 gene occurs following UVB irradiation. Cultured keratinocytes were exposed to UVB (290-320 nm, 300 J per m2) and then incubated at 37 degrees C for various intervals before harvesting. Immediately following UV exposure, the heat shock transcription factor 1 (HSF1) dissociated from HSP72-HSF1 complexes, underwent trimerization and phosphorylation, and demonstrated DNA binding activity to the heat shock element in the promoter region of the hsp72 gene. UVB also increased hsp72 mRNA, with peak levels observed 1-3 h post-UVR. HSP72 protein was constitutively expressed in keratinocytes, and its expression was increased by UVB, with maximum levels at 6 h post-UVR. The stress response may be extremely important in the protection of human skin from UVB radiation, and modulation of heat shock protein expression and/or function offers a potential therapeutic target in the prevention of photoaging and skin cancer.
Article
The 27 kDa heat shock protein (HSP 27) is expressed in keratinocytes of the upper epidermal layers, and recent evidence suggests that this protein is involved in the regulation of epidermal differentiation. The expression of HSP 27 was investigated in developing human skin by immunohistochemistry utilizing a specific monoclonal antibody. We used formalin-fixed, paraffin-embedded tissue of abdominal skin obtained from 34 human fetuses ranging between 13 and 30 weeks estimated gestational age (EGA). We found that HSP 27 is not expressed in keratinocytes until week 14 EGA. At this stage staining is observed in the periderm and the upper intermediate cells but not in hair germs. During further development, HSP 27 expression correlates with increasing epidermal differentiation, i.e. shedding of the periderm and beginning of keratinization. HSP 27 expression is confined to the upper cell layers and sparse basal cells. In hair follicles, HSP 27 can be detected in the innermost cell layer of the outer root sheath and in keratinocytes of the bulge identical to what is observed in adult skin. The hair papilla, matrix cells and sebaceous glands are negative for HSP 27 and remain so during further development. In eccrine sweat glands of the 24th week EGA, HSP 27 is confined to the superficial cell layer of the sweat ducts. In the present report we demonstrate differentiation-related expression of HSP 27 in developing human skin. Further in vitro studies will address the molecular function of HSP 27 in epidermal differentiation and development.
Article
Heat shock proteins (HSPs) stabilize intracellular processes of cells under stress. Little is known about the role of HSPs in wound healing, or whether their expression is altered by systemic disease. The focus of this study was to examine the local heat shock response to wounding in diabetic mice. Congenitally diabetic and phenotypically normal mice underwent standardized full-thickness cutaneous wounding. Mice were sacrificed at sequential time points and the wound beds excised. Tissues underwent immunohistochemical (IHC) and RT-PCR analyses for inducible HSP70. HSP70 protein expression in the wound bed by IHC peaked at 24 h in the nondiabetic mice. Expression of HSP70 was delayed in the diabetic mice until Day 3, which correlates with the clinical delay in healing seen in this model. The protein was especially prominent in the epithelium and in inflammatory cells migrating into the granulation tissue matrix. RT-PCR demonstrated upregulation of HSP70 mRNA within 12 h after wounding, lasting until Day 3, and decreasing thereafter in both the nondiabetic and the diabetic animals. Cutaneous wounding produces a HSP response in inflammatory cells, and expression of inducible HSP70 is delayed in diabetic mice. This delay may be related to the impaired inflammatory response of diabetics, and may contribute to impaired wound healing. The wound may be a continuing source of the heat shock response in inflammatory cells after injury.
Article
This study was performed to test the hypothesis that expression of heat shock proteins (HSPs) exhibits a spatially selective response within intact human skin following in vivo exposure to thermal stress. This response is believed to protect cells and tissues from further damage. Using Western blotting and immunohistochemistry, we studied the expression of a range of HSPs in normal human skin of 5 subjects prior to and following heating in vivo. The skin was heated to 41 +/- 0.5 degrees C for 1 h and biopsies were taken at 4, 8 and 24 h and from control, untreated skin. HSPs 27, 60, 72i, 90, 110 and heat shock constitutive (HSC)70 were expressed in normal skin, but the extent and distribution of these HSPs showed considerable variation. HSP27, 60 and 72i were found predominantly in the epidermis, whereas HSC70 showed weak epidermal staining but strong dermal expression. Heating the skin in vivo resulted in an increased skin content of HSP27, 60, 72i and 90, with maximal increase at 24 h following hyperthermia, while the skin content of HSC70 and HSP110 were unchanged. Significant increases in the content of HSP72i and HSP90 had occurred by 4 h following hyperthermia, with a mean +/-SEM of 206 +/- 50% and 197 +/- 38% of the control, untreated values, respectively (p<0.05). These findings indicate the complexity of HSP dynamics in human skin, and suggest that heating within the experimental range may protect the skin from further stresses for at least 24 h.
Article
sHsp (small stress proteins) are molecular chaperones involved in cellular defence mechanisms against several different types of aggressions. These proteins also participate in essential physiological processes, such as regulation of cell cycle, differentiation, programmed cell death and tumorigenicity. For example, sHsp are transiently expressed during the cell division to differentiation transition and this phenomenon prevents differentiating cells from undergoing apoptosis. sHsp also protect against apoptosis induced by different conditions or agents, particularly anti-cancer drugs. Of interest, tumor cells usually express high levels of sHsp, and anti-cancer drugs, such as cisplatin, trigger the accumulation of sHsp. These proteins are also known to interfere with programmed cell death induced by TNF alpha and Fas ligand. Moreover, they enhance the growth of tumors in vivo. Taken together, these observations suggest that sHsp can allow cancerous cells to escape the immunosurveillance mediated by death ligands and can render these cells resistant to therapy. Hence, sHsp represent prime targets for therapeutic interventions. This review is focused on the role of sHsp in different aspects of the life and death of mammalian cells and on the role of these survival proteins in cancer.
Article
Human papillomavirus type 16 (HPV16) infection has been linked to the development of cervical and anal dysplasia and cancer. One hallmark of persistent infection is the synthesis of the viral E7 protein in cervical epithelial cells. The expression of E7 in dysplastic and transformed cells and its recognition by the immune system as a foreign antigen make it an ideal target for immunotherapy. Utilizing the E7-expressing murine tumour cell line, TC-1, as a model of cervical carcinoma, an immunotherapy based on the administration of an adjuvant-free fusion protein comprising Mycobacterium bovis BCG heat shock protein (hsp)65 linked to HPV16 E7 (hspE7) has been developed. The data show that prophylactic immunization with hspE7 protects mice against challenge with TC-1 cells and that these tumour-free animals are also protected against re-challenge with TC-1 cells. In addition, therapeutic immunization with hspE7 induces regression of palpable tumours, confers protection against tumour re-challenge and is associated with long-term survival (> 253 days). In vitro analyses indicated that immunization with hspE7 leads to the induction of a Th1-like cell-mediated immune response based on the pattern of secreted cytokines and the presence of cytolytic activity following antigenic recall. In vivo studies using mice with targeted mutations in CD8 or MHC class II or depleted of CD8 or CD4 lymphocyte subsets demonstrate that tumour regression following therapeutic hspE7 immunization is CD8-dependent and CD4-independent. These studies extend previous observations on the induction of cytotoxic T lymphocytes by hsp fusion proteins and are consistent with the clinical application of hspE7 as an immunotherapy for human cervical and anal dysplasia and cancer.
Article
Normal-mode ruby laser (NMRL) irradiation of skin has now become an acceptable method of producing depilation. However, side effects, which include superficial burning and changes in skin pigmentation, still occur and, although temporary, can be distressing to the patient. This paper reports a method by which the skin can be protected (or preconditioned) from damage during NMRL treatment by pre-heating to a lower, non-damaging level prior to irradiation. Using the black-haired mouse (C57B1/10) as an animal model, an appropriate 'preconditioning' temperature was established by exposing the mouse skin to a range of temperatures, taking biopsies and staining the skin immunohistochemically for heat shock protein 70 (HSP 70) expression within the keratinocyte cells. Increased HSP 70 expression is stimulated by exposure to environmental stressors such as heat, so it was hypothesised that its increased expression conveyed increased cellular protection. The appropriate temperature (45 degrees C for 15 min) allowed for the superficial skin cells to be protected (as assessed by maximal HSP 70 staining) but undamaged (as assessed by haematoxylin and eosin staining), leaving the target hair-producing cells unprotected. Eight mice (16 flanks) were then exposed to this preconditioning temperature (eight of the flanks being growing-hair sites and eight resting-hair sites) and 5 h later exposed to a laser fluence known to cause mild skin damage and depilation (6J/cm2). This exposure was to both the preconditioned and the adjacent non-preconditioned sites. A statistically significant reduction in skin damage (P <0.001), as measured by the time taken to heal and noted both clinically and histologically, was seen in the preconditioned sites in resting-hair regions but not in growing-hair regions. Depilation, established over an 8 week period, was successful in growing-hair regions within both preconditioned and non-preconditioned sites, but complete hair regrowth had occurred in preconditioned and non-preconditioned sites within resting-hair regions by 5 weeks. The latter finding was consistent with work already reported suggesting that NMRL-assisted depilation in this animal model is not successful for hairs in the telogen phase. Successful preconditioning of mouse skin prior to laser exposure appears to reduce NMRL-induced skin side effects. In addition, the technique does not appear to adversely affect successful depilation.
Article
We previously reported that skin closure is improved by photoirradiation of the wound margins with an 815-nm diode laser system. To determine whether the beneficial effects of laser treatment involve the overexpression of the inducible 72-kDa heat shock protein, Hsp70. Expression of Hsp70 was investigated by immunocytochemistry in normal hairless rat dorsal skin and compared with its expression after laser photoirradiation. Constitutive expression of Hsp70 was mainly confined to the upper epidermal layer. Laser irradiation further increased epidermal expression of Hsp70 while inducing de novo synthesis of the protein in dermal structures, particularly around blood vessels, hair follicles and sebaceous glands. Laser-induced expression of Hsp70 was still present 7 days after photoirradiation. Laser-induced expression of Hsp70 might contribute to improved tissue regeneration and wound healing.
Specific induction of 70kDa HSPs by Herbimycin-A protects cardiomyocytes: a pharmacological route to cytoprotection
  • Morris
Small stress proteins as novel regulators of apoptosis
  • Mehlen